%0 Journal Article
%T Construction of Expression Plasmids Harbouring Genes Encoding Recombinant FN Polypeptides with Triple-domain and Preliminary Characterization of the Products Expressed in Escherichia coli<br>三结构域重组FN多肽表达质粒的构建及其表达产物性质的初步鉴定
%A LI Ming-Cai
%A FENG Zuo-Hua
%A LI Dong
%A ZANG Gui-Mei
%A <br>李明才
%A 冯作化
%A 李东
%A 张桂梅
%J 生物工程学报
%D 2000
%I 
%X To investigate the important role of recombinant triple-domain FN polypeptide in tumor therapy, two expression plasmids pF94-62 and pF94-82 were constructed and used to express triple-domain polypeptides of human FN in E. coli. The expressed polypeptides were CH62 (Pro 1239-Ser 1515 of FN linked with Ala 1690-Val 2049 through Met) and CH82 (CH62 without Pro 1953-Glu 1978). CH82 polypeptide was expressed as inclusion bodies in E. coli cultured at 37 degrees C. After denaturation with 8 mol/L urea and renaturation, the polypeptides were purified by the affinity chromatograph with Heparin-agarose, and the purified product was analysed by cell adhesion assay. The expression level of CH62 in E. coli was very low(5%), but that of CH82 was very high (21%), it suggested that N terminal sequence of Cell II in FN was the key sequence which influence the expression of triple-domain polypeptide in E. coli. The purified product was capable of binding heparin and cells, and it had a better binding activity than bifunctional-domain FN polypeptides. The production of CH82 polypeptide provided a fundamental basis for further study of recombinant product with better function of anti-metastasis and immune regulation.
%K Fibronectin
%K recombinant polypeptide
%K metastasis<br>纤连蛋白
%K 重组多肽
%K 肿瘤转移
%K 三结构域
%K 肿瘤治疗
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=B4EF61671A4FAD7E&yid=9806D0D4EAA9BED3&vid=7801E6FC5AE9020C&iid=E158A972A605785F&sid=FE6B7E9BDCCDBAA6&eid=F18BA6286A889C1C&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=1&reference_num=5