Expression,indentification and functional characterization of RC-RNase
Jun Zhao · Zhonghua zhongliu fangzhi zazhi · 2005
OBJECTIVE:TO express RC-RNase in E.coli Bl21(DE3)plysS and to observe its killing effect on Hepatocellular carcinoma cell in vitro. METHODS: Prokaryctic expression vetor containing PET32a-RC-RNase was induced to be expressed by IPTG in E.coli Bl21(DE3)plysS.The E.coli was lysed by ultrasonication.Deposit of the lysate was purified by Ni-NTA Agarose affinity chromatography. Use MTT method to detect the killing effect on Hepatocellular carcinoma cell SMMC-7721 in vitro. RESULTS: There was a new anticipated band on SDS-PAGE gel.The form of the expressed protein was container.The results of cytotoxic assay showed that the targeting protein could kill hepatocarcinoma cell and its IC_ 50 is 22.44 μg/mL. CONCLUSION: The satisfied effect would be the base to research its others functions.