Published Date: Jun 01 2024
Recombinant plasmid is a commonly used technique in recombinant DNA technology. In the process of recombinant plasmid construction, a series of recombinant enzymes are needed to complete the various operations. In this article, we will introduce the recombinant enzymes and their roles in the recombinant DNA technology process.
Restriction Endonucleases
Restriction endonuclease is one of the basic recombinant enzymes. It recognizes a specific DNA sequence of a DNA molecule and cuts the double-stranded DNA at this column. The choice of restriction endonuclease is very important, considering the DNA sequence's characteristics and the operations to be performed. Commonly used restriction endonucleases are EcoRI, BamHI, HindI, etc.
Ligase
Ligase plays a role in joining different DNA segments in recombinant plasmid construction. Ligases catalyze the breaking and synthesis of phosphodiester bonds in the DNA molecule to join the sticky ends of two DNA fragments. One of the most commonly used ligases is T4DNA ligase.
Filler Enzymes
The role of filler enzymes in recombinant plasmid construction is to reduce the sticky ends produced by enzymatic cleavage to form an uncut circular DNA molecule. Common filler enzymes include T4DNA polymerase, Klenow enzyme, etc.
Splitting, Ligating, And Modifying Enzymes
These recombinant enzymes perform split ligation and modification of specific DNA fragments during recombinant plasmid construction. They can cut DNA molecules at specific sites in the target DNA sequence or ligate and modify them after cutting. Common splitting, ligating, and modifying enzymes include Exonuclease, Polynucleotide Kinase, etc.
Recombinant plasmid construction is one of the commonly used technical tools in genetic engineering, in which recombinant enzymes play a crucial role. The selection and use of these recombinant enzymes must be determined according to the experimental needs and the characteristics of the target genes. Through the rational selection and use of recombinant enzymes, recombinant plasmid construction can be carried out efficiently, supporting subsequent gene expression studies.
