柔嫩艾美耳球虫甘肃株钙调域蛋白激酶基因的克隆与表达  

Cloning and Expression of Calmodulin-domain Protein Kinases Gene (CDPK)of Eimeria tenella Gansu Strain

作　　者：周建民[1] 鲍杰 汪明[1] 

机构地区：[1]中国农业大学动物医学院,北京100094 [2]北京市兽医卫生监督所,北京100044

出　　处：《畜牧兽医学报》2005年第7期711-714,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金："863"基金项目(2002AA245061)

摘　　要：应用反转录聚合酶链式反应(RTPCR)技术,从柔嫩艾美耳球虫甘肃株(E.tenellaGS,EtGS)孢子化卵囊的子孢子中提取总RNA扩增得到鸡球虫子孢子表面抗原钙调域蛋白激酶(CDPK)基因。将EtGSCDPK基因与原核表达载体pGEX6P1连接,构建了pGEXCDPK原核表达质粒,并获得高效表达和纯化的CDPK融合蛋白,表达率达35.4%。序列分析表明:EtGSCDPK与文献报道的EtCDPK比较,共有5个核苷酸发生变异,核苷酸同源性为99.6%;有5个氨基酸发生变异,氨基酸同源性为98%。The calmodulin-domain protein kinases(CDPK)gene was cloned by RT-PCR from total RNA which was extracted from sporozoite of Eimeria tenella Gansu strain. Using the CDPK gene, the pGEX-CDPK recombinant plasmid was constructed. The recombinant plasmid was transformed into E.coli BL21 and highly expressed. The acquired fusion protein was purified by glutathione resin volume.The amount of fusion protein in total bacteria protein were 35.4%. The CDPK gene of Eimeria tenella Gansu strain was consisted of 1 470 nucleotides, encoding 490 amino acids. Compared with the CDPK gene of Eimeria tenella that reported in GenBank there are five mutation nucleotides and the homology of nucleotides is 99.6%, and there are five mutation amino acids and the homology of CDPK amino acids is 98%.

关 键 词：鸡球虫 CDPK 克隆 表达 

分 类 号：S852.722[农业科学—基础兽医学]