中文摘要：

为研究柔嫩艾美耳球虫（Eimeria tenella）细胞色素bc 1复合物（cytochrome bc1,Cytbc1）的生化特性,本研究对其基因进行了克隆和原核表达。根据EupathDB已公布的E.tenella Houghton株Cytbc1基因序列（Contig ID：ETH 00007855）设计特异性引物,运用RT-PCR方法扩增EtCytbc1基因序列,并将其克隆到表达载体pCold43a中,构建重组质粒pCold43a-EtCytbc1,经测序鉴定正确后,将其转化感受态细胞E.coli Rosetta（DE3）,并进行IPTG诱导表达。结果表明,EtCytbc1的ORF序列（687bp）,编码228个氨基酸;重组菌pCold43a-EtCytbc1能成功诱导表达,pCold43a表达部分可溶性蛋白,表达产物纯化后可用于酶生化特性分析。柔嫩艾美耳球虫Cytbc1基因的成功克隆表达为该酶的功能研究奠定了基础。

英文摘要：

In order to analyze biochemical properties of E.tenella cytochrome bc1（EtCytbc1）,the full-length of open reading frame of cytochrome bc1 was amplified by RT-PCR from total RNA from the second-generation schizonts of E.tenella.And then cloned into pCold43 avector.This plasmid was transformed into E.coli Rosetta（DE3）and induced by IPTG.The expression products were analyzed by SDS-PAGE and then purified.The results showed that the Cytbc1 gene consists of a 687 bp open reading frame encoding 228 amino acids.Sequencing analysis showed that the recombinant vector pCold43a-EtCytbc1 was constructed successfully.The SDS-PAGE results showed that the recombinant proteins were partially soluble.Soluble protein was obtained from pCold43aEtCytbc1,and was purifide by the affinity chromatography assay.The purified protein can be used for biochemical analysis of this enzyme.The prokaryotic expression of EtCytbc1 provides a basis on the molecular characterization of E.tenella cytochrome bc1.