Question

A researcher studying llama genetics would like to clone her gene of interest into the pUC19 cloning vector using the blue-white screening approach we discussed in class. Below are the vector and llama DNA fragment to be cloned into the vector. The restriction enzyme sites indicated on both DNA molecules. Which restriction enzyme should she use in order to get the largest possible DNA fragment cloned into the vector?

Answer 1

She should use type II restriction enzymes  in order to get the largest possible DNA fragment cloned into the vector.

Type II restriction enzymes are the familiar ones used for everyday molecular biology applications such as gene cloning and DNA fragmentation and analysis.

These enzymes cleave DNA at fixed positions with respect to their recognition sequence, creating reproducible fragments and distinct gel electrophoresis patterns.

Over 3,500 Type II enzymes have been discovered and characterized, recognizing some 350 different DNA sequences. Thousands more ‘putative’ Type II enzymes have been identified by analysis of sequenced bacterial and archaeal genomes, but remain uncharacterized.

Type II restriction enzymes are very diverse in terms of amino acid sequence, size, domain organization, subunit composition, co-factor requirements and modes of action.

They are loosely classified into a dozen or so sub-types according to their enzymatic behaviour. This is a practical classification that reflects their properties rather than their phylogeny.

It does not necessarily reflect evolutionary or structural relationships, and the subtypes are not mutually exclusive. An enzyme can belong to several subtypes if it exhibits each of their defining characteristics.

Learn more about restriction enzymes here : brainly.com/question/15278286

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