Direct cloning of the newest angiogenesis inhibitor,arresten / 重庆医科大学学报

Objective:To clone and express the new angiogenesis inhibitor,arresten.Methods:The total RNA was extracted from the normal human liver,and then was reversely transcribed mRNA to cDNA and arresten cDNA was amplified by nestle-PCR.After being treated by T4 DNA polymerase,the arresten cDNA was linked with the linear vector pTYB1 digested by NdeI and EcoRI.And then the complete ER2566 was transfected by recombinant vector,the recombination was screened and the expression of the interest protein was induced by IPTG.Results:The arresten was cloned by one-step cloning method with PCR product treated by T4 DNA polymerase.Conclusion:This method can be used to clone the interest gene more easily and quickly than T-A clone and others.