1. An unusual plasmid has been discovered which has high transformation ability in E.coli and confers heat resistance to the host. Hence the host E.coli when transformed with this plasmid can grow at 50 degree celsius. Schematically represent the various steps of coning a foreign gene in this plasmid. How would you screen the transformed E.coli carrying this plasmid?
2.Given the sequence of the two primers (5 nucleotide long)required to amplify the following DNA sequence by PCR: 5'GCACCTAGATCGATCC3'
3. You wish to introduce the human insulin gene into a bacterial host in the hope of producing large amounts of human insulin. Should you use genomicDNA or cDNA? Explain.
4. Explain why the sequence read from the autoradiogram is complementary to the original sequence?
5. What kind of promoters should be used in expression vectors?
6. How will you select yeast cells transformed with Leu2 gene?
7. Viral vectors are ideal vehicles for transfer of foreign DNA into host cell. Why? Give an example of both plant and animalviral vectors.
8. What are the properties of major enzymes required in dideoxychain termination method?
9. Explain the process of automated DNA sequencing when you got the following sequence from its autoradiogram 5'AGCTATCAG3'
10. Listed below are two different sets of ssDNA of their ds form which one would you expect it to be cleaved by a R.E. and why?
a. 3'AGCCTTCCGGATGCGC5'
b. 3'GCTTAAGCTTTCTGCAA3'
2.Given the sequence of the two primers (5 nucleotide long)required to amplify the following DNA sequence by PCR: 5'GCACCTAGATCGATCC3'
3. You wish to introduce the human insulin gene into a bacterial host in the hope of producing large amounts of human insulin. Should you use genomicDNA or cDNA? Explain.
4. Explain why the sequence read from the autoradiogram is complementary to the original sequence?
5. What kind of promoters should be used in expression vectors?
6. How will you select yeast cells transformed with Leu2 gene?
7. Viral vectors are ideal vehicles for transfer of foreign DNA into host cell. Why? Give an example of both plant and animalviral vectors.
8. What are the properties of major enzymes required in dideoxychain termination method?
9. Explain the process of automated DNA sequencing when you got the following sequence from its autoradiogram 5'AGCTATCAG3'
10. Listed below are two different sets of ssDNA of their ds form which one would you expect it to be cleaved by a R.E. and why?
a. 3'AGCCTTCCGGATGCGC5'
b. 3'GCTTAAGCTTTCTGCAA3'
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