Transformation:

Process in biotechnology to introduce foreign DNA into a host organism.

Alters host cell membrane permeability to uptake foreign DNA.

Fundamental technique in genetic engineering for creating genetically modified organisms (GMOs).

Discovery of Transformation:

Discovered by Frederick Griffith in 1928.

Observed heat-killed pathogenic bacteria could transform non-pathogenic bacteria into a pathogenic form.

Pivotal in understanding genetic material transfer.

Competent Cells:

Host cells treated to enhance foreign DNA uptake.

Made more permeable through chemical treatments or electroporation.

Allows efficient introduction of recombinant DNA.

Screening of Recombinant Clones:

Identifying host cells with incorporated recombinant DNA.

Methods include antibiotic resistance markers, reporter genes, and DNA sequencing.

Ensures selection of cells with desired genetic modifications.

Choices of Host and Vectors:

Selection of host organism and vector (plasmid or viral DNA) is critical.

Different organisms and vectors have unique traits for specific applications.

E. coli commonly used for molecular cloning due to rapid growth and well-understood genetics.

Microbial Growth and Protein Production:

Microbial growth essential for protein production.

Cultivation of host organisms under controlled conditions (nutrients, temperature, aeration).

Leads to expression of recombinant proteins for medical and industrial applications.

Protein Production at Industrial Scale:

Crucial for meeting demands in pharmaceuticals and biomanufacturing.

Bioreactors (large fermentation vessels) used to cultivate microbial cells for efficient and economical protein production.

Downstream Processing:

Follows microbial growth and protein production.

Includes purification, separation, and isolation of target protein from cellular components.

Techniques like chromatography, filtration, and centrifugation used for obtaining pure, functional proteins.