The Ti (tumor-inducing) plasmid is a large plasmid found in the bacterium Agrobacterium tumefaciens. This plasmid is responsible for transferring a segment of its DNA into plant cells, causing the formation of crown gall tumors. The Ti plasmid is widely utilized in plant genetic engineering to introduce new genes into plants.
Researchers exploit the Ti plasmid's natural ability to transfer DNA to manipulate plant genomes. By replacing the tumor-inducing genes with desired genes, scientists can create genetically modified plants with beneficial traits, such as pest resistance or increased yield.
Answer the following questions:
(a) [(i)] Explain how some strains of Bacillus thuringiensis produce proteins that kill certain insects such as lepidopterans but do not kill the Bacillus.
[(ii)] How is the above mechanism exploited for the production of Bt cotton plant by biotechnologists?
(b) [(i)] Explain how the amplification of gene of interest is done using PCR.
[(ii)] State two applications of the desired amplified fragment of DNA.
The basic scheme of the essential steps involved in the process of recombinant DNA technology is summarised below in the form of a flow diagram. Study the given flow diagram and answer the questions that follow.
(a) What is the technical term used for Step 4 in the above process?
(b) Which of the given two combinations of restriction enzyme should be used in Step 1? Justify your answer.
(i) EcoR I to cut the plasmid and Hind III to cut the alien DNA.
(ii) EcoR I to cut both the plasmid and alien DNA.