链激酶基因的分离与表达及其单克隆抗体的制备(英文)  

Isolation and Expression of Streptokinase Gene and Preparation of Its Monoclonal Antibody

作　　者：周俐梅 王嘉玺[1] 王利红 王静[1] 邹民吉 赵春文 段聚宝 

机构地区：[1]北京基础医学研究所,北京100850

出　　处：《中国实验血液学杂志》1996年第4期387-393,共7页Journal of Experimental Hematology

摘　　要：用PCR方法从链球菌染色体基因组中分离了链激酶基因,并把它克隆到表达载体pBV220中。诱导表达后重组蛋白以包涵体的形式存在并占菌体总蛋白的60％以上。纯化后的重组蛋白的纯度达90％以上。血纤维蛋白原平板测活方法表明,重组蛋白比活性为1.3×10~5U/mg。重组链激酶蛋白免疫小鼠后获得6株分泌抗链激酶单克隆抗体的杂交瘤。把重组链激酶基因克隆至M13mp19中进行DNA序列分析,结果表明与文献报道的有区别。We amplified recombinant streptokinase (rSK) gene by PCR from the chromosomal DNA of Streptococus pvogenes group A and expressed it in the expression vector pBV220, and the expressed rSK amounted to 60% of total celi protein. The final purity of the rSK reached above 95% after purification by gel filtration and anion - exchange chromatography. The specific activity was 1. 3 × 105 U/mg determined by tbe fibrinogen plate technique. Six hybridoma clbnes which secreted monoclonal antibodies against rSK were obtained. The rSK gene was cloned into the vector M13 mp19 for DNA sequencing. The nucleotide sequenee more or less differs from the previously reported and it shares 91% homology with another SK gene SPSKA isolated from another strain of Streptococcus pyogenes, and 87% homology with SK gene SESKG from Streptococcus equsimilis group C.

关 键 词：链激酶 链激酶基因 血纤维蛋白溶酶原 基因表达 单克隆抗体 

分 类 号：R346[医药卫生—基础医学]