细粒棘球蚴热休克蛋白70基因的克隆和序列分析

目的 获得细粒棘球蚴（E.granulosus）热休克蛋白（Heat Shock Protein70，HSP70）基因，进行DNA测序、序列特性分析及同源性分析，为研究包虫病重组疫苗候选基因奠定基础。方法 从细粒棘球蚴原头蚴提取总RNA，RT－PCR技术扩增出细粒棘球蚴中国大陆侏HSP70基因，将其重组到pGEM－T载体后进行序列测定和分析。结果 成功扩增出细粒棘球蚴中国大陆侏HSP70基因，测序表明该片段由402bp组成，与已发表基因核苷酸序列相比，同源性为96％，推导编码氨基酸序列同源性为81％。结论 经DNA测序证实，扩增出的片段确为HSP70，该片段阅读框完整，可作为重组抗原的候选基因。

Cloning and sequence analyzing of the HSP70 gene from echinococcus granulosus

Objective To obtain and analyze characters and homologies of sequence HSP70 gene, and lay bases for screening candidate antigen of echinococcus granulosus. Methods Total RNA was extracted from protoscoles of cysts. The HSP70 gene of ecbinococcus granulosus was amplified by RT- PCR and recombined into pGEM- T vector for sequencing and analyzing. Results A cDNA sequence with an open reading frame of 402bp has been amplified successfully by RT- PCR. Compared with the DNA sequence published, the hcmologies are 96%, which deduced that the amino acid sequence of HSP70 of echinococcus granulosus are 81% identities. Conclusion By analyzing the DNA sequence,we can make sure that the segment of amplification is HSP70, and its reading frame is integrity. As a result, it can he used as the potential gene of recombination antigen.