Cloning, Sequencing of Suaeda heteroptera kitag CMO cDNA and Construction of its Recombinant Plant Expression Vector / 中国生物工程杂志

Total RNA was extracted from leaf of Suaeda hetroptera kitag, then the CMO ( choline monooxygenase) cDNA was amplified using the reverse transcriptase polymerase chain reaction ( RT-PCR) method and cloned into pMD-T-simple vector. The positive clones from the Blue/White Screen were sequenced. After confirming its validity, the CMO gene fragment was cloned into pBI121 vector. Double enzyme restriction and PCR analysis indicated that the pBI121/CMO recombinant plasmid was successfully constructed.

Expression in E.coli JM109 and pharmacology on anti-angiogenesis of endogenetic angiogenesis inhibitor Arresten gene / 中国药理学通报

Aim To construct prokaryotic expression vector for endogenetic angiogenesis inhibitor Arresten gene and to express recombinant.Pharmacology experiments found that the expression product had the function of restrain blood vessel growing on the chorioallantoic membrane(CAM).Methods The total RNA was extracted from the placenta organize of the normal puerpera,and Arresten gene was amplified by RT-PCR to construct recombinant vector pBV220-Arr.Then,it was transformed into E.coli JM109 for yielding recombinant human Arresten protein.The method to detect inhibiting angiogenesis activity was CAM study.Results The plasmid pBV220-Arr.could be expressed in E.coli JM109 from two hours to eight hours,and the expression yield reached the highest in two hours.The Arresten protein could restrain blood vessel growth of CAM evidently than that of angiostatin.Conclusion The recombinant plasmid pBV220-Arr was construted successfully and the target protein Arresten can express in E.coli JM109,the protein of Arresten had the evident effect of inhibite the activity of angiogenesis.