2014•Guocheng gongcheng xuebaoRequires access
Expression and Anti-agiogenesis Activity of Human and Mouse Endostatin
Yang Yi-ji
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Abstract
Human endostatin(HE) and mouse endostatin(ME) cDNA sequences were determined based on the reports and NCBI database. Their 554 bp HE and 565 bp ME cDNA sequences were obtained through DNA synthesis, respectively. Prokaryotic expression plasmids pGEX-4T1-HE and pET-32a-ME with different protein labels of GST and TrxA were constructed, then expressed through induction in Escherichia coli BL21(DE3), HE and ME recombinant proteins were obtained and their anti-angiogenesis activity was analyzed by chick chorioallantoic membrane(CAM) test. The human and mouse endostatin gene expression vectors with different labels were successfully constructed and used in induced expression in Escherichia coli. CAM testing results indicated that the purified refolded recombinant protein had a strong anti-angiogenesis activity. The human and mouse ES recombinant proteins were obtained successfully. TrxA and GST labels showed no difference in the increase of soluble expression of recombinant ES, both of the recombinant protein with different labels mainly existed in inclusion body.