Hello, everyone welcome you back on the topic: human genome project and DNA fingerprinting. So, starting with the human genome project, Human Genome project was a mega project that aimed to sequence every base in Human Genome.

Human Genome project was a mega project in which various countries, so many peoples, were involved to find out the bases present in the human genome. This project has helded much new information, so many new information was given by Human Genome project. Many new areas and avenues have opened up as a consequences of the project because of human genome projected so many different new areas was being opened up so that the further experimentation can be done in Human Genome. This project began in 1990, 15 years effort. 18 countries participated in the worldwide effort, so you can see it is not a small project. It was a mega project in which almost 18 countries were involved and it took almost 15 years to being completed.

In September 99, there was a public announcement about this human genome project and a rough draft y draft. By spring 2000, a draft was published in year two thousand about ninety percent of the genome. Human genome was being completed and a complete high quality DNA sequence was given in 2003, so it was started in 1990 and was completed by 2003.. Human genome project assembles 12 000 bases.

Every minute you can imagine in a single minute. 12 000 bases was assembled by this project. 15 billion raw base pairs were sequenced to reach the two billions, each area of chromosome, at least four, to five times to ensure that the data deposited is accurate. So whenever certain data was being deposited as a raw information, it was checked by four to five times to ensure the accuracy of the project and there was a depth of coverage. So many different sequences were given by human genome project goals of human genome project. First was to identify all the exp, approximately twenty thousand, to twenty five thousand genes in human DNA. Earlier before human genome project. It was expected that more than thirty five thousand genes are there in human being, but h g p concluded that approximately in human being there are twenty thousand to twenty five thousand genes present in human DNA determine the sequences of three billions chemical base pair that make Up human DNA, so, according to HGP in human beings, there is approximately three billions base pairs. Those information can be stored as databases in computer, whatever databases, whatever information was gathered in human genome project that was stored as database in the computer for future experimentation.

Human genome project improve tools for data analysis, whatever data was gathered that can be analyzed on various parameter so that they can be utilized, transfer related technologies to other sectors such as industries. Whatever information was gathered that can be given to another industry for their experimentation, and it also addresses the ethical, legal and social issues that may arise from the project. There are so many ethical, legal and social issues that was answered by human genome project. There are some solid features of human genome project.

The human genome contains three thousand one hundred sixty four point: seven millions, nucleotide bases the average gene consists of 3000 bases, but sizes vary greatly with the largest known human gene being dystrophin at 2.4 million bases. This information was given by human genome project. The total number of gene is estimated at 30 000 much lower than previous estimated of eighty thousand to one lakh.

Forty thousand genes, almost all nucleotide bases, are exactly the same in all living organism. So ninety nine point: nine percent base sequences are similar in all the living organisms in case of human being, so the all human beings are 99.9 percent genetically similar.

The function are unknown for over 50 percent of discovered genes, even though the genes were discovered, but the HGP given the answers of those unknown genes by giving their functions less than two percent of genome codes for protein. So I called h g p states that less than 2 percent of whole genome of human being codes for the protein, that means less than 2 percent of human genome is functional.

More than 98 percent is non-functional, repeated sequences may up make up very large portion of the human genome. A human genome have very large number of repetitive sequences. Repeated sequences are stretches of DNA sequences that are repeated many times sometimes hundred to thousand times. They are thought to have no direct coding function, but they shed light on chromosome structure dynamics and evolution. So there are so many repetitive sequences present in DNA that can be repeated many times and that may be hundred two thousand times.

But these repeated sequence do not have any function to pro to synthesize a protein, but they have certain function related to chromosome structure, its dynamic and evolution. Chromosome one has most gene. If we go for karyotyping. As we know, kerotyping is arrangement of chromosome according to their length, so the first chromosome has the most genes. The number of genes is much more in the first number of chromosome. They have 2968 genes in their chromosome, and y chromosome has the fewest only 231 genes. Are there in y chromosome scientists have identified about 1.4 millions location where single based DNA differences - SNP single nucleotide polymorphism occurs in humans. So, with the help of this HGP scientist identify that about 1.4 millions, location, 1.4 milli location have s and p, which makes us very much specific.

When we, when we talk about DNA finger painting, then this information promises the revolutionize the process of finding chromosomal location for disease associated sequences and trusting human history. So these information was very much important to find out any type of genetic disorder which leads to the disease. In any human being benefits of HGP research, if we talk about benefits, the first area is medical benefits. In case of medical, there is improved diagnosis of disease. Disease diagnosis has been improved with the help of information given by the HGP earlier detection of predisposition of disease. With the help of this HGP, the earlier damage, mutation or reason of disease can be detected.

Rational, drug design and the drug can be designed according to the disease. Gene therapy and control system for drug gene therapy is correcting the wrong nucleotide or mutation, either by replacing the gene or by salinity the gene pharmacogenomics personal drugs. Various type of drugs pharmaceuticals has been designed for various disease and organ replacement.

The information given by h g p also helps in organ transplantation. Another area of h, g p research is microbial genome research, new energy sources. As biofuels, the microbial cells can be used to get biofuels. Environmental monitoring to detect pollutants pollutants can be detected using the information given by HGP protection from biological and chemical warfare, safe, efficient, toxic waste cleanup. Ah, as we know today, we are facing the problems of pollution with the various toxic chemicals that can be cleaned up by using the microbial genome, which was given by HGP in case of forensic science. The HGP research help us to identify potential suspects at crime scenes. The criminals can easily be identified which it will help us to identify the criminals. We will learn in DNA, finger printing, exonerate, wrongly accused person. H g p also helps to rescue to rescue the wrong one. The person who has not performed any crime and was arrested for certain reason. It also helps to identify crime and catastrophe victims. It establishes the paternity and other family relations the paternity dispute. The child dispute was solved by the HGP research, identify endangered and protected species.

As an aid to wildfire officials, so many endangered species of animals have been identified and their genome has been preserved, detect bacteria and other microorganisms that may pollute air water, soil and food. So the bacteria which causes the pollution of air water, soil and food has been detected and there in disinfection has been given by the HGP, match organ donors with recipient in transplant programs during transplantation. As we know, we need to match the foreign organs with the recipient.

This matching was easily been done by the HGP research determine pedigree for seed or livestock breeds authenticate consumables such as caviar and wine. So these are certain benefits of h, g p, research in case of agriculture and livestock disease. Insect and drought resistant crop has been created, healthier, more productive disease resistance forms, animals has been produced, more nutritious plants has been produced, bio pesticides have been produced, edible vaccines incorporated into food products was given by HGP research.

Do you know what is adival vaccine? Edible vaccines? Are those vaccines which are taken as food in the form of fruits or any part of the plant new environmental cleanup uses for plant like tobacco? So many different researches is also being done in that field of agriculture and livestock in case of evolution and human immigration? It uses germline mutation in linear, stress story, evolution study, migration of different population groups based on female genetic inheritance study mutation on the evolutionary, stable y chromosome to truss, lineage and migration. Compare break points in the evolution of mutation with edges of population and historical events, assess health damage and risk caused by the radiation exposure, including low dose exposure. So if we are being exposed by certain radiation that may cause the mutation in the DNA and that can be assessed by the human genome, project, assess health damage and risk caused by exposure to mutagenic, chemicals and cancer-causing toxic chemicals. So HGP also helps to identify various type of mutation caused by chemical mutagen, reduce the likelihood and heritable mutation. So HGP was a mega project which gives us so many information and still the research are going on so that it can be exploited for better experimentation.

Now the next topic of today’s session is DNA fingerprinting. Earlier, the coming of the biotechnology in the society, the finger printing was being used to identify the criminals. Every individual have a specific finger printing pattern and accordingly, they would be identified nowadays this fingerprinting are done in DNA. So what is DNA? Fingerprinting? DNA fingerprinting is a technique to find out variation in individual of a population at DNA level or DNA. Fingerprinting is a method used to identify an individual from a sample of DNA by looking at unique pattern in their DNA, so DNA finger printing help us to identify the variation in individual in any living organism by looking up into the DNA, as we know, we all Are 99.9 percent genetically similar, only 0.1 to 0.2 percent? We are genetically dissimilar, and this differences in DNA makes us very much a specific in our genome and because of this changes in DNA, we are. We have a specific DNA in our genome, which will leads to identification of various criminals even in case of child dispute, paternity dispute or in case of rape. It works on the principle of polymorphism in DNA sequences, so DNA fingerprinting works on polymorphism of DNA polymorphism means variation, so many different sequences are there in our DNA in our genome and that polymorphism that variation makes us genetically specific than that of another human being.

Although we are 99.9 percent genetically similar, but because of this polymorphism in that 0.1 to 0.2 percent of DNA makes sets genetically a specific, and because of that, we will be identify easily with the help of DNA finger printing here, some backgrounds of DNA fingerprinting, almost every Cell in our body contains our DNA, as we know, we are made up of cells and every cells contain.

The DNA on average about 99.9 percent of DNA between two humans is same. So if we talk about similarity in human beings, you can see. U and me is not very much similar. We have different patterns, we have different phenotype, but if we talk about a gene level, we are 99.9 percent genetically similar.

The remaining percentage is what makes us unique. Remaining percent means. 0.1 percent of DNA is dissimilar in between you and me, and that differences makes us very much specific. Although phenotypically we are dissimilar, but genetically we are 99.9 percent similar. Only the 0.1 percent dissimilarity in genes makes us specific. You are specific because of that 0.1 percent, and i am specific because of 0.1 percent of that genome, although this might should sound like a small amount, it means that there are around three millions base pair that are different between two people.

These differences can be compared and used to help distinguish from one one from someone else. So, according to h g p, we have approximately 3 into 10 to the power 9 base pairs and these 3 into 10 to the power 9 base. Pairs have some differences, which leads to the 0.1 percent dissimilarity and makes us genetically specific. Many satellites are short sequences, approximately 10 to 16 base pairs of repetitive DNA that show greater variation from one person to next then other parts of the genome.

This variation is exhibited in the numbers of repeated units of or stutters in, the mini satellite sequences, so many different type of polymorphism in DNA, one is mini satellite, which is also considered as repetitive sequences, which are being repeated again and again in the genome. The length of that DNA fragment may be from 10 to 16 base pairs.

The first mini satellite was discovered in 1980. DNA finger printing was invented in 1984 by professor sir, alex afraid after he realized you could have detect variation in human DNA in the form of this mini satellite. So DNA fingerprinting was given by professor sir alex zafre in eighty four DNA finger. Printing is a technique that simultaneously detects lots of mini satellite in the genome to produce a pattern unique to an individual earlier. The mini satellite was first utilized by the sir alex zephyr referred to identify a particular unknown human being nowadays. So many different type of poly form is polymorphism is there which is being utilized in DNA fingerprinting.

The probability of having two people with the same DNA fingerprints that are not identical. Twins are very small, so two people having the same DNA finger printings is rare to happen living accepting the twins. The twins have the exact DNA copies so leaving the twins accepting the twins. No human beings can have same DNA fingerprints. Just like your actual fingerprints. Your DNA fingerprint is something you are born with. It is a unique to you so as we have a unique pattern of fingerprinting pattern in our fingers, like that, every human beings have a unique DNA finger. Printings now here are some steps which is being followed in DNA finger printing. The first is isolation of DNA. Suppose we need to go for daily finger printing. So first you have to select a cell from this cells. We can isolate the DNA, so first step of DNA finger. Printing is isolation of DNA. This DNA will be isolated from the cell.

Next step is digestion of DNA by restriction in the nucleus. As we know, this DNA is very much bulky in nature. This DNA will be fragmentized by using restriction. Endonuclease enzyme restriction to nucleus enzyme is an enzyme which cleaves the DNA, and it is also known as molecular seizure. Why molecular scissor means cutter, so it cut the DNA. That is why it is considered as molecular seizure when this DNA is mixed with restriction to nuclease. It will fragmentize this DNA in different fragments of different length like this.

The next step after the cleavage by restriction on nucleus is separation of DNA fragments by electrophoresis. Now all these fragments these are fragments of DNA. Now this these fragments of DNA will be separated on gel electrophoresis.

When these fragments are transported transferred on a gel electrophoretic apparatus, these fragments will be separated according to their length. Do you know why DNA can move under the influence of electricity to move to travel under the influence of electricity? A chemical compound must contain certain charges, and DNA also contain that charge. Overall, there is negative charge on the DNA because of phosphate group, so because of that negative charge, this DNA can move under the influence of electricity and when it undergo gel electrophoretic apparatus, all these fragments will be separated out after that, gel electrophoretic photic apparatus is like This here we have a gel tank. This is gel tank. The gel may be of agarose or polyacrylamide. Here we have electricity, cathode and anode. Here we have buffer tank.

Now these fragments of DNA is loaded on this gel and the electricity is being passed on because of that electricity. These fragments will move inside the gel. Ah, nylon membrane is put over this gel so that the DNA can go and stick into the nylon or nitrocellulose membrane after that, after the separation by gel electrophoresis, the nylon sheet is being taken out, and then these nylon seat or nitrous nitrocellulose membrane.

There is probe hybridization probe, hybridization means probe is artificially synthesized oligonucleotides, which may be radio, labeled or fluorescent level. So, these probes will hybridize the DNA present in this nylon or nitrocellulose membrane, and then this membrane will undergo autoradiography after autoradiography. You will find the pattern of DNA on the membrane, the bands you can see the bands of DNA and this band of DNA on the membrane is called auto radio gram and now these autoradiogram is being compared by different patient different peoples, different criminals, to identify an Unknown person, so I hope the steps would be clear to you. The first will take the cell from where we have to isolate the DNA will isolate the DNA being bulky in nature. The DNA will be fragmentized into different fragments of different lengths, using restriction and nuclease. These fragments will be separated according to their length by using gel electrophorus. Photonic apparatus will put a nylon or nitrocellulose membrane over the gel. All the fragments, separated fragments of DNA will stick on. This membrane will take out the membrane. The transfer of DNA from gel to membrane is called blotting, and the whole technique is also considered, as also known as southern blotting. It was given by a scientist southern. That is why it is known as southern blotting and transfer of DNA from gel to membrane is called blotting. Now this nylon or nitrocellulose membrane has DNA, this DNA will be hybridized with the probe is artificially synthesized oligonucleotide. Now, wherever the probe will be hybridized in the DNA, the other probes which have not hybridized, will be removed off by the washing and after that, the nylon will undergo autoradiography after rotography will find the pattern of DNA on the membrane, and this pattern of DNA is Called autoradiogram, which is being matched by the different peoples. So, these are the steps of DNA fingerprinting. You can take the blood sample. As a cell you will isolate.

The DNA from the blood cells will fragmentize the DNA we are using. The restriction nucleus will transfer the DNA fragments and separate out all DNA fragments on the gel electrophoretic apparatus. You will put a nylon or nitrocellulose membrane over the gel so that the DNA can stick on. The membrane will take out. The membrane will go for the probe. Hybridization, after probe hybridization, the nylon will undergo autoradiography by using the x-ray.

And finally, you will get this pattern of DNA on the membrane, and this pattern of DNA is being matched by different people. Different persons, DNA source from where we can isolate the DNA.

What are the different sources of DNA? The sources of the DNA may be blood, maybe semen may be vaginal. Fluid may be hair roots, skin skeletal or remains or somewhere else. So, these are certain sources of DNA from where we can isolate the DNA for DNA. Fingerprinting molecular markers are very important aspects of DNA. Fingerprinting, as we know, we are 99.9 percent, similar only 0.1 percent. We are genetically dissimilar and these 0.1 percent DNA have molecular markers, which makes us specific molecular markers are of different types. It may be RFLP that is, restriction, fragment length, polymorphism VNTR, variable number tandem, repeats short tandem, repeats str, SNP single nucleotide polymorphism RAPD random amplified polymorphic DNA. So these are certain molecular markers, which makes us very much specific, and because of this, only we will be identify if we are doing any crime. Here are some applications of DNA fingerprinting. The first application is in forensic science.

The second application is paternity. If any dispute is related to any child or any parent, that dispute can easily be solved using the DNA finger printing genetic biodiversity, biodiversity, genetically genetics can be easily be studied. Using DNA, fingerprinting and evolutionary biology evolution can easily be studied using DNA finger printing. What are the differences in evolution we can easily detect using DNA finger printing. Now let us answer. This question spends HGP, yes, human genome project, HGP. How many base pairs are there in human? According to HGP, we have approximately 3 into 10 to the power 9 base pairs. What is DNA fingerprinting DNA finger printing is a technique. Is a method to identify the variation in the individual among the population so that the disputes can be solved? What is the principle of DNA? Fingerprinting DNA finger. Printings works on polymorphism in the DNA. Many of the DNA fragments show polymorphism that polymorphism in the DNA is called molecular marker that may be RAPD RFLP STR, VNTR, SNP and etcetera mention some sources from where DNA can be isolated from DNA finger printing for DNA. Fingerprinting from where we can isolate the DNA. We can isolate the DNA from the blood from the semen, vaginal fluid root, hairs and etcetera. So, I hope this question must be clear and the special session would be useful for us, so I hope the HGP and DNA fingerprinting must be clear to you and you would be able to answer the question which is there in your textbook before any optimization. Let’s answer the question which I asked in my earlier session and the question which was asked to my student. The first question is: what is gene expression so which we learn in regulation of gene expression. Gene expression means how a gene express itself. That is how a DNA transfer is information to RNA and then from RNA to protein. So, formation of protein from DNA via RNA is called gene expression in case of RNA. As we know, there are three types of RNAs, mainly transfer, RNA, messenger, RNA and ribosomal RNA. So out of these three RNA, mainly the messenger RNA codes for protein, but ribosomal RNA and transfer RNA indirectly help in protein synthesis. So how a gene the gene that is functional part of the DNA which will synthesize the protein so how a DNA synthesized protein through RNA is called gene expression. The transfer of genetic information from DNA to RNA is called transcription and transfer of information from RNA to protein is called translation. So we can conclude over here. Gene expression is transcription plus translation, so transcription along bin translation is called gene expression formation of protein through DNA via RNA. Next question is: what do you mean by operon? If you talk about prokaryotic cells, prokaryote cells have an operon system to control their gene expression controlling of gene expression means whenever any protein or enzymes are required by the cell. In that condition, the gene should express itself to synthesize the protein, and whenever we do not need any type of protein or enzymes, the gene should not express itself. It should be in switch off condition. So, in the case of prokaryotes, there are a group of genes which work together to control the expression of gene that group of genes which control the expression of genes is called operon. The operon consists of regulatory genes operator, promoter and structural genes. All these genes combine together to form an operon system and the operon system is called the regulatory sequences of the prokaryotic cell. The next question is differentiated between inducible and repressible operon. This we learn in regulation of gene expression in prokaryote in case of prokaryotes. Can you differentiate prokaryotes from eukaryotes? Yes, prokaryotes do not have a nuclear membrane to enclose the genetic material as well as the prokaryotes lacks the membrane bounded organelle like mitochondria, like chloroplast, like endoplasmic, reticulum, golgi, body and etcetera. So, in case of prokaryotes, the operon system is there, and this operon system help the prokaryotic cell, especially the bacteria to control the expression of gene, and this operon is of two types in case of bacteria. One is inducible and another is repressible in case of inducible. The genes are normally switched off in case of inducible operon system. Normally the genes are switched off means. The genes are not expressing themselves to synthesize the protein, but the genes can be induced to express itself. So the genes which are in switch of condition normally can be induced by certain factor. Certain substrate to express themself, an example of inducible operon system is lac. Operon lac means lactose. Lactose is one of the carbohydrates which gives the energy to the living system. In case of repressible operon system, the genes are normally switched on in case of repressible operon, the normally the genes are expressing themselves to synthesize the protein and they are in switched on condition and to repress to switch off the such type of genes. The genes are repressed to stop gene expression. So here you can see. Example, we have loc lac apparent in case of inducible operon system and in case of repressible the example is TRP operon system. So, we can conclude the differences in inducible operand system. The genes are normally switched off, while in case of repressible system the genes are normally switched on. So, if the genes are normally switched off, they have to make certain proteins or enzymes whenever they are required. So, genes are made to induce to synthesize the protein to express themselves, but in case of repressible operand system, the genes are normally switched on, so they are being repressed whenever we do not need such type of protein or amino acid, and then in that condition the Genes are repressed to stop. The gene expression example of inducible operon system is lac. The operon and repressible operon system is TRP operon, so I hope the differences must be clear to you. Inducible and repressible operon system, which is found in prokaryotic cell, especially in bacteria, give an example of irritable and repressible operon system example of inducible operon system is lac, operon and repressible operon system is TRP, operon, lac, coperon means lactosaperone and TRP means tryptophan operon system. If you can recall the concept, the lacaperone have gene organization, like this lag said like y and black x. Here they have operator and then promoter and at the some distance they have lag. I repressor gene and the promoter of the repressor gene. So, this lac, I synthesize the repressor protein. Normally this repressor protein is active. It can bind over the promoter and will block the RNA polymerase to move along the structural genes, but in the presence of lactose some of the lactose is converted into allolactose. These allolactose will interact with this these repressor protein and will inactivate it, and because of that, these repressure protein, which is there over the operator, will be removed and the operator will be free. The pro the at the promoter, the RNA polymerase, can bind and will transcribe these genes to synthesize the protein. So this is lac operon lactose Perone, which is an example of inducible operon system, while in case of TRP operon TRP operon is repressible, operon and TRP means tryptophan. Tryptophan is one of the amino acid. If we talk about the organization of DNA, there are five genes, TRPa TRPb, TRPc, TRPd and TRPe with the operator and promoter. Here there is TRP, I repressor protein will be synthesized. The repressor protein is normally inactive. It cannot bind over the operator and the transcription translation would be there, but in the presence of tryptophan this protein will be inactive, protein will be activated and it will be bind over. The operator blocks operator blocks RNA polymerase to move among the structural genes. No transcription translation would be there and the gene expression can be switched off, so this is an example of TRP operon system that is repressible operand system. The next question is what is monosystronic and polycystronic DNA. In case of bacteria, you can see for the synthesis of tryptophan these genes. Are there TRP a b c d e, like in the case of lac apparent? There are three genes lag, z, lac, y and lac x, lag z, will synthesize b tag, electro side is lack y, will synthesize permease and lag x will synthesize acetyl transferase. So, all these genes are expressed to synthesize different genes, different proteins, but in case of eukaryotes, these genes are separated from each other. There are certain non-functional sequences between the two genes, but in the case of bacteria or prokaryotes, these genes are close together. They are situated together and they are expressed simultaneously once the beta galactose had been synthesized. At the same time, the permias enzyme will be synthesized and next to that, acetyl transverse enzyme will be synthesized, so these genes are present together are considered as polycystronic. In the case of bacteria, but in the case of eukaryotes, the genes are present at some distance there is a gap between two genes, so one gene will express at a certain time and in different time another gene will be expressed. So, such type of DNA is called Mono systemic DNA, which is present in eukaryotes next question, which was asked by my students how thymine dimers are formed in case of DNA. You know if it is a DNA here it is a nucleotide, nucleotide, nucleotide. Here also in the next strand, these are the nucleotides and there it is a hydrogen bond in between the two strands, and this bond is called phosphor diastere bond in between the two nucleotides. This is the phosphodiester bond. Phosphodastrobond is created between the sugar of one nucleotide and the phosphate of another nucleotide. There is no any bond in between the nitrogenous bases. Nitrogenous bases are bonded by hydrogen bond between two strands, but if any, how suppose? Here we have thymine nucleotide, and here also thymine nucleotide is there, and there is bond formation unusual bond formation between the nitrogenous bases in a strand which is unusual. This type of bond formation is called thymine, thymine dimer. This is one of the type of mutation, and this is caused by a raise which is called as UV rays, that is ultraviolet rays. So ultraviolet rays causes the thymine dimer formation in a strands of the DNA which cause the mutation next question: why UV rays cause the mutation you can see? UV rays can cause a mutation because UV rays provides lost lots of energy and because of that extra energy, there will be unusual formation of bond. The next question is differentiate between repetitive, repetitive DNA and satellite DNA. Repetitive, DNA is the DNA sequences which are being repeated again and again in a genome. While satellite DNA is one of the one of the type of repetitive DNA which then, when separated, they will form a satellite pathway, a ring pathway, which is same as that of the satellite. That’s why it is considered as satellite DNA. So here I am going to end my session hope you have enjoyed a lot.