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Question
Identify the option with the correct match for I, II and III.
Identify the option with the correct match for I, II and III.
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Solution
The correct option is B I - lacZ, II - Selectable marker, III - Selects recombinants
The lacZ gene is present as a selectable marker in many cloning vectors to screen recombinants from non-recombinants. The gene codes for 𝛃-galactosidase enzyme. This enzyme hydrolyses the chromogenic substrate, X-gal, into a blue product.
- The colonies of bacteria that have taken up the recombinant vector, having the gene of interest inserted within the lacZ gene, will appear white due to insertional inactivation of the lacZ gene.
- Those bacterial colonies transformed with the non-recombinant vector that lacks the insert (gene of interest) within the lacZ gene will appear blue due to the functional lacZ gene.
EcoR I is a restriction enzyme which recognizes the following palindromic sequence in double stranded DNA and cleaves it between G and A. It creates staggered / sticky ends and not blunt ends.
5' —— GAATTC —— 3'
3' —— CTTAAG —— 5'
YAC is Yeast artificial chromosome which is a linear DNA as it is obtained from yeast, an eukaryote. It is used to clone very large DNA fragments.
Plant cells are bombarded with high velocity micro-particles of gold or tungsten coated with DNA in a method known as biolistics or gene gun. It is a direct method of gene delivery as the host cell is directly getting transformed.
The lacZ gene is present as a selectable marker in many cloning vectors to screen recombinants from non-recombinants. The gene codes for 𝛃-galactosidase enzyme. This enzyme hydrolyses the chromogenic substrate, X-gal, into a blue product.
- The colonies of bacteria that have taken up the recombinant vector, having the gene of interest inserted within the lacZ gene, will appear white due to insertional inactivation of the lacZ gene.
- Those bacterial colonies transformed with the non-recombinant vector that lacks the insert (gene of interest) within the lacZ gene will appear blue due to the functional lacZ gene.
EcoR I is a restriction enzyme which recognizes the following palindromic sequence in double stranded DNA and cleaves it between G and A. It creates staggered / sticky ends and not blunt ends.
5' —— GAATTC —— 3'
3' —— CTTAAG —— 5'
YAC is Yeast artificial chromosome which is a linear DNA as it is obtained from yeast, an eukaryote. It is used to clone very large DNA fragments.
Plant cells are bombarded with high velocity micro-particles of gold or tungsten coated with DNA in a method known as biolistics or gene gun. It is a direct method of gene delivery as the host cell is directly getting transformed.
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