Question

Before a desired gene can be spliced into a bacterial plasmid cloning vector, the gene must first be ________ from the original DNA

Answer 1

Before splicing a gene into a bacterial plasmid, it must be isolated from original DNA, typically using a restriction enzyme to cut the DNA, then ligated into the plasmid.

Step-by-step explanation:

Before a desired gene can be spliced into a bacterial plasmid cloning vector, the gene must first be isolated from the original DNA. This process often involves several steps:

• The target DNA from a donor organism is extracted and cleaved enzymatically by a selected restriction endonuclease.
• The plasmid is removed from a bacterium, and its DNA is also cut by the same restriction enzyme to ensure that both the plasmid and the foreign DNA can be joined together compatibly.
• Then, the foreign DNA and the plasmid are ligated together by the enzyme DNA ligase, creating a new recombinant DNA molecule known as a cloning vector-insert DNA construct.
• Finally, the recombinant plasmid can be introduced into a bacterial host for replication and expression of the desired gene.

The process of isolating and inserting DNA into a plasmid vector is a fundamental technique in genetic engineering and biotechnology that allows researchers to study and utilize genes in a bacterial system.