<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Intoduction</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Biotechnology deals with techniques of using live organisms or enzymes from organisms to produce products and processes useful to humans</p> </li> <li> <p>In this sense, making curd, bread or wine, which are all microbe-mediated processes, could also be thought as a form of biotechnology</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px"><span style="color:blue">Intoduction</span> $\rarr$ Principles of Biotechnology $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Introduction</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>However, it is used in a restricted sense today, to refer to such of those processes which use genetically modified organisms to achieve the same on a larger scale</p> </li> <li> <p>Further, many other processes/techniques are also included under biotechnology</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px"><span style="color:blue">Intoduction</span> $\rarr$ Principles of Biotechnology $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Introduction</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>For example, in vitro fertilisation leading to a ‘test-tube’ baby, synthesising a gene and using it, developing a DNA vaccine or correcting a defective gene, are all part of biotechnology</p> </li> <li> <p>The European Federation of Biotechnology (EFB) has given a definition of biotechnology that encompasses both traditional view and modern molecular biotechnology</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px"><span style="color:blue">Intoduction</span> $\rarr$ Principles of Biotechnology $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Introduction</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li>The definition given by EFB is as follows: ‘The integration of natural science and organisms, cells, parts thereof, and molecular analogues for products and services’</li> </ul> </div> </main> <hr> <footer style= "font-size:18px"><span style="color:blue">Intoduction</span> $\rarr$ Principles of Biotechnology $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li>Among many, the two core techniques that enabled birth of modern biotechnology are :</li> </ul> <p>(i) Genetic engineering : Techniques to alter the chemistry of genetic material (DNA and RNA), to introduce these into host organisms and thus change the phenotype of the host organism</p> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>(ii) Bioprocess engineering : Maintenance of sterile (microbial contamination-free) ambience in chemical engineering processes to enable growth of only the desired microbe/eukaryotic cell in large quantities for the manufacture of biotechnological products like antibiotics, vaccines, enzymes, etc</p> </li> <li> <p>Let us now understand the conceptual development of the principles of genetic engineering</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>You probably appreciate the advantages of sexual reproduction over asexual reproduction</p> </li> <li> <p>The former provides opportunities for variations and formulation of unique combinations of genetic setup, some of which may be beneficial to the organism as well as the population</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Asexual reproduction preserves the genetic information, while sexual reproduction permits variation</p> </li> <li> <p>Traditional hybridisation procedures used in plant and animal breeding, very often lead to inclusion and multiplication of undesirable genes along with the desired genes</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>The techniques of genetic engineering which include creation of recombinant DNA, use of gene cloning and gene transfer, overcome this limitation and allows us to isolate and introduce only one or a set of desirable genes without introducing undesirable genes into the target organism</p> </li> <li> <p>Do you know the likely fate of a piece of DNA, which is somehow transferred into an alien organism? Most likely, this piece of DNA would not be able to multiply itself in the progeny cells of the organism</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>But, when it gets integrated into the genome of the recipient, it may multiply and be inherited along with the host DNA</p> </li> <li> <p>This is because the alien piece of DNA has become part of a chromosome, which has the ability to replicate</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>In a chromosome there is a specific DNA sequence called the origin of replication, which is responsible for initiating replication</p> </li> <li> <p>Therefore, for the multiplication of any alien piece of DNA in an organism it needs to be a part of a chromosome(s) which has a specific sequence known as ‘origin of replication’</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Thus, an alien DNA is linked with the origin of replication, so that, this alien piece of DNA can replicate and multiply itself in the host organism</p> </li> <li> <p>This can also be called as cloning or making multiple identical copies of any template DNA</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Let us now focus on the first instance of the construction of an artificial recombinant DNA molecule</p> </li> <li> <p>The construction of the first recombinant DNA emerged from the possibility of linking a gene encoding antibiotic resistance with a native plasmid (autonomously replicating circular extra-chromosomal DNA) of Salmonella typhimurium</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Stanley Cohen and Herbert Boyer accomplished this in 1972 by isolating the antibiotic resistance gene by cutting out a piece of DNA from a plasmid which was responsible for conferring antibiotic resistance</p> </li> <li> <p>The cutting of DNA at specific locations became possible with the discovery of the so-called ‘molecular scissors’– restriction enzymes</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>The cut piece of DNA was then linked with the plasmid DNA</p> </li> <li> <p>These plasmid DNA act as vectors to transfer the piece of DNA attached to it</p> </li> <li> <p>You probably know that mosquito acts as an insect vector to transfer the malarial parasite into human body</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>In the same way, a plasmid can be used as vector to deliver an alien piece of DNA into the host organism</p> </li> <li> <p>The linking of antibiotic resistance gene with the plasmid vector became possible with the enzyme DNA ligase, which acts on cut DNA molecules and joins their ends</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>This makes a new combination of circular autonomously replicating DNA created in vitro and is known as recombinant DNA</p> </li> <li> <p>When this DNA is transferred into Escherichia coli, a bacterium closely related to Salmonella, it could replicate using the new host’s DNA polymerase enzyme and make multiple copies</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>The ability to multiply copies of antibiotic resistance gene in E. coli was called cloning of antibiotic resistance gene in E. coli</p> </li> <li> <p>You can hence infer that there are three basic steps in genetically modifying an organism —</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Principles of Biotechnology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <p>(i) identification of DNA with desirable genes;</p> <p>(ii) introduction of the identified DNA into the host;</p> <p>(iii) maintenance of introduced DNA in the host and transfer of the DNA to its progeny</p> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ <span style="color:blue">Principles of Biotechnology</span> $\rarr$ Tools of Recombinant DNA Technology $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Now we know from the foregoing discussion that genetic engineering or recombinant DNA technology can be accomplished only if we have the key tools, i.e., restriction enzymes, polymerase enzymes, ligases, vectors and the host organism</p> </li> <li> <p>Let us try to understand some of these in detail</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p><strong>Restriction Enzymes</strong></p> </li> <li> <p>In the year 1963, the two enzymes responsible for restricting the growth of bacteriophage in Escherichia coli were isolated</p> </li> <li> <p>One of these added methyl groups to DNA, while the other cut DNA</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>The later was called restriction endonuclease</p> </li> <li> <p>The first restriction endonuclease–Hind II, whose functioning depended on a specific DNA nucleotide sequence was isolated and characterised five years later</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>It was found that Hind II always cut DNA molecules at a particular point by recognising a specific sequence of six base pairs</p> </li> <li> <p>This specific base sequence is known as the recognition sequence for Hind II</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Besides Hind II, today we know more than 900 restriction enzymes that have been isolated from over 230 strains of bacteria each of which recognise different recognition sequences</p> </li> <li> <p>The convention for naming these enzymes is the first letter of the name comes from the genus and the second two letters come from the species of the prokaryotic cell from which they were isolated, e.g., EcoRI comes from Escherichia coli RY 13</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>In EcoRI, the letter ‘R’ is derived from the name of strain</p> </li> <li> <p>Roman numbers following the names indicate the order in which the enzymes were isolated from that strain of bacteria</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Restriction enzymes belong to a larger class of enzymes called nucleases</p> </li> <li> <p>These are of two kinds; exonucleases and endonucleases</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Exonucleases remove nucleotides from the ends of the DNA whereas, endonucleases make cuts at specific positions within the DNA</p> </li> <li> <p>Each restriction endonuclease functions by ‘inspecting’ the length of a DNA sequence</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li>Once it finds its specific recognition sequence, it will bind to the DNA and cut each of the two strands of the double helix at specific points in their sugar -phosphate backbones (Figure 9.1)</li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <img src= "https://temp-public-img-folder.s3.ap-south-1.amazonaws.com/sathee.prutor.images/images/ncertbook/bio/b12/biotechnology/ncert_b12_ch09_endonuclease_enzyme_EcoRI.png"> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Each restriction endonuclease recognises a specific palindromic nucleotide sequences in the DNA</p> </li> <li> <p>Do you know what palindromes are? These are groups of letters that form the same words when read both forward and backward, e.g., “MALAYALAM”</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>As against a word-palindrome where the same word is read in both directions, the palindrome in DNA is a sequence of base pairs that reads same on the two strands when orientation of reading is kept the same</p> </li> <li> <p>For example, the following sequences reads the same on the two strands in 5’$\rarr$ 3’ direction</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>This is also true if read in the 3’$\rarr$ 5’ direction Restriction enzymes cut the strand of DNA a little away from the centre of the palindrome sites, but between the same two bases on the opposite strands</p> </li> <li> <p>This leaves single stranded portions at the ends</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li>There are overhanging stretches called sticky ends on each strand (Figure 9.1)</li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <img src= "https://temp-public-img-folder.s3.ap-south-1.amazonaws.com/sathee.prutor.images/images/ncertbook/bio/b12/biotechnology/ncert_b12_ch09_endonuclease_enzyme_EcoRI.png"> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>These are named so because they form hydrogen bonds with their complementary cut counterparts</p> </li> <li> <p>This stickiness of the ends facilitates the action of the enzyme DNA ligase</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Restriction endonucleases are used in genetic engineering to form ‘recombinant’ molecules of DNA, which are composed of DNA from different sources/genomes</p> </li> <li> <p>When cut by the same restriction enzyme, the resultant DNA fragments have the same kind of ‘sticky-ends’ and, these can be joined together (end-to-end) using DNA ligases (Figure 9.2)</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <img src= "https://temp-public-img-folder.s3.ap-south-1.amazonaws.com/sathee.prutor.images/images/ncertbook/bio/b12/biotechnology/ncert_b12_ch09_recombinant_dna_technology.png"> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>You may have realised that normally, unless one cuts the vector and the source DNA with the same restriction enzyme, the recombinant vector molecule cannot be created</p> </li> <li> <p>Separation and isolation of DNA fragments : The cutting of DNA by restriction endonucleases results in the fragments of DNA</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>These fragments can be separated by a technique known as gel electrophoresis</p> </li> <li> <p>Since DNA fragments are negatively charged molecules they can be separated by forcing them to move towards the anode under an electric field through a medium/matrix</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Nowadays the most commonly used matrix is agarose which is a natural polymer extracted from sea weeds</p> </li> <li> <p>The DNA fragments separate (resolve) according to their size through sieving effect provided by the agarose gel</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>Hence, the smaller the fragment size, the farther it moves</p> </li> <li> <p>Look at the Figure 9.3 and guess at which end of the gel the sample was loaded</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <img src= "https://temp-public-img-folder.s3.ap-south-1.amazonaws.com/sathee.prutor.images/images/ncertbook/bio/b12/biotechnology/ncert_b12_ch09_electrophoresis_showing_migration.png"> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>The separated DNA fragments can be visualised only after staining the DNA with a compound known as ethidium bromide followed by exposure to UV radiation (you cannot see pure DNA fragments in the visible light and without staining)</p> </li> <li> <p>You can see bright orange coloured bands of DNA in a ethidium bromide stained gel exposed to UV light (Figure 9.3)</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <img src= "https://temp-public-img-folder.s3.ap-south-1.amazonaws.com/sathee.prutor.images/images/ncertbook/bio/b12/biotechnology/ncert_b12_ch09_electrophoresis_showing_migration.png"> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>

<p><Success style = "float: center; text-align: center; font-size:30px"><B>Biotechnology : Principles and Processes</B></Success></p> <p><Primary style = "float: center; text-align: center; font-size:26px"><B>Tools of Recombinant DNA Technology</B></Primary></p> <hr> <main> <div style='float: left; width:100%; text-align: left;font-size:23px'> <ul> <li> <p>The separated bands of DNA are cut out from the agarose gel and extracted from the gel piece</p> </li> <li> <p>This step is known as elution</p> </li> <li> <p>The DNA fragments purified in this way are used in constructing recombinant DNA by joining them with cloning vectors</p> </li> </ul> </div> </main> <hr> <footer style= "font-size:18px">Intoduction $\rarr$ Principles of Biotechnology $\rarr$ <span style="color:blue">Tools of Recombinant DNA Technology</span> $\rarr$ Processes of Recombinant DNA Technology</footer>