pET28TEV-AapCas12b Expression Plasmid

The pET28TEV-AacCas12b plasmid is the expression plasmid of AacCas12b (also known as AacC2c1) protein, which is used to express and prepare AacCas12b protein in Escherichia coli BL21 (DE3) strain. AacCas12b nuclease is an RNA-mediated endonuclease that can specifically cut target double-stranded DNA in the presence of PAM (TTN) in the target DNA. The AacCas12b protein is smaller than Cas9 and Cas12a, and is derived from Alicyclobacillus acidoterrestris, an acidophilic thermostable bacterium with high cleavage activity. The optimal cleavage reaction temperature is 48°C. Similar to Cas12a of the same family, it recognizes the PAM sequence of 5'-TTN and cleaves double-stranded DNA to generate cohesive ends. AacCas12b is guided by dual RNA, so it relies on crRNA and tracrRNA (or sgRNA formed after ligation). In addition, AacCas12b can not only be applied to the cleavage of target dsDNA, but its trans-cleavage activity against ssDNA can also be used for rapid detection of target nucleic acids. The plasmid contains a lactose operon inducible expression system, which can be induced to express the target protein by IPTG or lactose and its analogues.

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