Gene cloning,expression and preliminary activity analysis of hBcl-2 / 中国免疫学杂志

ABSTRACT

Objective:Through gene cloning,expression and activity analysis of hBcl 2 to provide enough proteins for the development of new drugs on the basis of 3 D structue of hBcl 2 protein.Methods:Gene cloning using PCR amplification,identification with Western Blot and MALDI TOF MS.Results:hBcl 2 gene was cloned,inserted into pET28a(+) and solublely expressed in E.Coli.Its molecule weight was confirmed through MALDI TOF MS,which fits exactly with its theoretical value.After purification to reach electrophoresis homogeneity,it showed the ability of combining specifically with BH 3 domain of Bak,and then provide the basis for the further research on small chemical compounds which could specifically bind hBcl 2 protein.Conclusion:In this work hBcl 2 was successfully expressed and recovered its binding activity in a soluble form. [