Synthesis and Expression of a Gene From Kringle-2 Domain of Tissue Plasminogen Activator in E. coli  

作　　者：华子春 傅和亮 陈于红 余瑞荣 王进 朱德煦 

机构地区：[1]Department of Biochemistry, Nanjing University, Nanjing 210008, PRC

出　　处：《Science China Chemistry》1994年第6期667-676,共10页中国科学（化学英文版）

基　　金：Project supported by a grant from the Chinese National Program of High Technology Development.

摘　　要：The DNA fragment corresponding to the tissue plasminogen activator (tPA) sequence 174-262 (Kringle-2 domain) has been synthesized by using the solid phase phosphotriester method. The Kringle-2 domain of human tPA was expressed in Escherichia colt by secretion into the periplasmic space using the Lpp-Lac promoter and PIN-Ⅲ OmpA2 signal sequence. About two thirds of the expression product was secreted into the periplasmic space , and purified with ammonium sulfate fractionation, affinity chro-matography on Lysine-Sepharose, and FPLC-Mono Q exchange chromatography. The amino acid composition observed from the Kringle-2 purified from E. coli is identical with that expected for the 174-262 fragment of human tPA. Radio binding assay shows that the recombinant Kringle-2 domain possesses the activity of fibrin binding.The DNA fragment corresponding to the tissue plasminogen activator (tPA) sequence 174-262 (Kringle-2 domain) has been synthesized by using the solid phase phosphotriester method. The Kringle-2 domain of human tPA was expressed in Escherichia colt by secretion into the periplasmic space using the Lpp-Lac promoter and PIN-Ⅲ OmpA2 signal sequence. About two thirds of the expression product was secreted into the periplasmic space , and purified with ammonium sulfate fractionation, affinity chro-matography on Lysine-Sepharose, and FPLC-Mono Q exchange chromatography. The amino acid composition observed from the Kringle-2 purified from E. coli is identical with that expected for the 174-262 fragment of human tPA. Radio binding assay shows that the recombinant Kringle-2 domain possesses the activity of fibrin binding.

关 键 词：tissue PLASMINOGEN ACTIVATOR (tPA) KRINGLE domain. 

分 类 号：R346[医药卫生—基础医学]