%0 Journal Article
%T Cloning,Soluble Expression and Mutant Activity Analysis of Lactate Dehydrogenase Gene from Plasmodium falciparum<br>恶性疟原虫乳酸脱氢酶基因克隆、可溶性表达及突变体活性分析
%A XU Xiao-Ling
%A YANG Rui-Yi
%A YANG Xue-Qin
%A FENG Li-Ling
%A ZENG Qing-Ping
%A <br>徐小玲
%A 杨瑞仪
%A 杨雪芹
%A 冯丽玲
%A 曾庆平
%J 生物工程学报
%D 2007
%I 
%X To establish a platform for high throughput screening and in vitro evaluating novel metabolic enzyme-targeted inhibitors towards anti-malarial drugs, a lactate dehydrogenase gene of Plasmodium falciparum (PfLDH) was amplified from the Hainan isolate FCC1/HN. The fusion expression vectors, pGEX-2TK and pET-29a( + ), were utilized to introduce the PfLDH gene into strains of Escherichia coli, BL21 and BL21 (DE3), for over-expression. Consequently, the enzymatic activity of PfLDH was successfully detected in the suspension of lytic bacteria. The PfLDH gene cloned in pGEX-2TK was mainly expressed as inclusion bodies, while the same gene cloned in pET-29a( + ) was nearly expressed in a soluble form of PfLDH, demonstrating the latter vehicle might be more suitable for the large-scale preparation of recombinant PfLDH. Furthermore, according to the electrophoregram of SDS-PAGE and the sequencing data, a series of truncated PfLDH sequences generated randomly from gene amplification were screened and cloned, from which four pre-matured genes with a terminator mutation, PfLDH-delta271, -delta236, -delta167 and -delta53 coding for 45, 80, 149 and 263 amino acid residues, were individually recovered. Through the gene expression and enzymatic activity measurement, the effect of pre-matured terminator mutation on the activity of PfLDH was evaluated, which should pave the way for probing the relationship between structure and function of PfLDH.
%K PfLDH<br>融合表达
%K 终止突变
%K 酶促活性
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=E8C2E3B03DD2140D&yid=A732AF04DDA03BB3&vid=EA389574707BDED3&iid=E158A972A605785F&sid=2EAE52BA5B1222A9&eid=0A8675156EB60B87&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=21