动物造模,长爪沙鼠CST3序列 z-bo 2020-09-25 324

　　OBJECTIVE: To analyze the cDNA sequence homology of Cystatin C (CST3) in Mongolian gerbils, establish a prokaryotic expression system for CST3 protein, and lay the foundation for the preparation of CST3 antibodies in Mongolian mussels and subsequent genetic function studies. Cloning of snake Cst3 cDNA sequence and homology analysis.

　　"Method: Digest and ligate the optimized sequence into the pET28a vector to complete the construction of the recombinant CST3 protein expression vector. The vector is transformed into competent cells, and the prokaryotic expression of CST3 protein is induced by isopropyl β-D-thiogalactoside (IPTG), SDS polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting (Western) Pass (verification) verification.

　　Result: The mouse Cst3 gene has low sequence homology with human and mouse Cst3 genes. The codon-optimized Cst3 expression sequence of snake was inserted into the pET28a plasmid to obtain the CST3 protein expression vector. Induction of 1 mmol/LIPTG for 12 hours at 37°C can produce a large amount of CST3 protein.

　　Conclusion: An in vitro expression system for the CST3 protein of Mongolian gerbils has been successfully constructed.