恶性疟原虫FCCl／HN株LDH基因的克隆及序列分析

目的 克隆并测定恶性疟原虫海南(FCCl／HN)株LDN基因序列，比较FCCl/HN株与国外分离株LDH基因序列的差异。方法 根据LDH基因已知序列设计合成一对引物，应用PCR技术从FCCl/HN株基因组DNA中扩增LDH基因，并将其克隆入pMDl8-T载体。用双脱氧链末端终止法进行基因序列测定，并用分子生物学软件进行不同分离株LDH基因序列的同源性比较。结果 PCR扩增得到特异的FCCl／HN株LDH基因片段并构建了pT—LDH重组质粒。恶性疟原虫FCCl／HN株LDH基因全长951bp，编码316个氨基酸。序列分析表明，我国恶性疟原虫FCCl／HN株与国外的3所、Honduras 1株LDH基因编码氨基酸序列完全一致。结论 成功克隆恶性疟原虫FCCl，HN株LDH基因。序列测定及同源性分析表明，FCCl/HN株与其它分离株的LDH基因序列高度保守。

Cloning and Sequence analysis of LDH gene of Plasmodium Falciparum isolate FCC1/HN

Objective To clone and determine the mucleotide sequences of the lactate dehydrogenase( LDH) gene of plasmodiu falciparum isolate FCC 1/HN, and analyze the differences of the sequences of LDH genes among isolate FCC 1IHN and other ones worldwide. Methods The LDH gene was amplified by PCR technique from genomic DNA of isolate FCC 1/HN. Then it was cloned into pMD-18T vector. The cloned LDH gene was deter- mined by the dideoxy chain termination method. And the molecular biology softwares were used to find out the dif- ferences of the sequences of the LDH genes among different P. fakipqrum isolates. Results The LDH gene of isolate FCC1/HN was specifically amplified, and the correct recombinant plasmid pT-LDH was contructed. The result of sequencing the nucleotides showed that the LDH gene of isolate FCC1/HN was 951 base pair in full length, encoding 316 aminoacids. The LDH genes of P. falcipantm among isolates FCC1/HN, 3D7 and Honduras 1 exhibited high homology. Conclusion The LDH gene of P. fa1ciparun isolate FCC 1/HN was successfully cloned and sequenced. Sequences analysis showed that the LDH genes of isolate FCC 1/HN and other ones were quite conserved.