BIOLOGY Write the short note.

1. Insulin is a peptide hormone produced by β -cells of islets of Langerhans of pancreas.
2. Insulin is essential for the control of blood sugar levels.
3. Disease Diabetes mellitus is caused due to inability to make insulin.
4. Insulin was discovered by Sir Edward Sharpey Schafer (1916) while studying Islets of Langerhans.
5. Hakura et al (1977) chemically synthesized DNA sequence of insulin for two chains A and B and separately inserted into two PBR322 plasmid vector.

Gilbert and Villokomaroff, 1978 produced insulin production using r-DNA technology.

1. The recombinant plasmids, containing insulin gene inserted by the side of β-galactosidase, were transferred into E. coli host.
2. The host produced penicillinase pnd pre-pro insulin.
3. Insulin is later separated by trypsin treatment.

1. Plasmids are small, extra-chromosomal, double stranded circular forms of DNA that replicate autonomously.
2. They are seen in bacterial cells, yeast and animal cell.
3. Plasmids are considered as replicons as they are capable of autonomous replication in suitable host.
4. The most commonly used vectors in r-DNA technology are plasmids as they replicate in E. coli.

Plasmid as a cloning vector should have a replication origin, a marker gene for antibiotic resistance, control elements like promoter, operator, ribosome binding site, etc. and a region where foreign DNA can be inserted. Naturally plasmids do not have all these features. Hence, they are constructed by inserting gene for antibiotic resistance. pBR 322, pBR320, paCYC177 are the constructed plasmids.Ti plasmid (for tumor-inducing) of Agrobacterium tumefaciens is an important vector for carrying new DNA in many plants. It contains a transposon, called T DNA, which inserts copies of itself into the chromosomes of infected plant cells. The transposon, with the new DNA, can be inserted into the host cell’s chromosomes. A plant cell containing this DNA, can then be grown in culture or induced to form a new, transgenic plant.

1. Restriction endonucleases have the ability to recognize specific sequences in DNA and cleave it.
2. They are 4 to 8 nucleotides long and characterized by a particular type of internal symmetry.
3. The specific site at which restriction endonuclease cuts the DNA is called recognition site or restriction site.
4. Each restriction endonuclease recognizes its specific recognition sequence.
5. Restriction cutting may result in DNA fragments with blunt ends or cohesive or sticky ends or staggered ends (having short, single stranded projections).

For example, recognition sequence of by the enzyme EcoRI is
3′ —– -CTTAA-G —–5′
5′ —– -G A A T T C —–3′
It is as palindrome, i.e. When read on opposite strand of DNA (3′ to 5′ or 5′ to 3′) it reads same.
When the enzyme EcoRI recognizes this sequence, it breaks each strand at the same site in the sequence i.e. between the A and G residues.