细粒棘球蚴中国大陆株诊断抗原P-29基因的克隆和序列分析

目的获得细粒棘球蚴（E.granulosus）诊断抗原（diagnostic antigen）P-29基因并进行序列分析。方法从包虫病患者体内获取细粒棘球蚴原头蚴提取总RNA，根据GenBank公共数据库检索出细粒棘球蚴诊断抗原P-29基因的已知序列设计一对引物，通过RT-PCR技术扩增出细粒棘球蚴中国大陆株诊断抗原P-29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株诊断抗原P-29基因，测序表明该片段由717bp组成，与已发表基因核苷酸序列相比，同源性为100％，推导编码氨基酸序列同源性为100％。结论成功克隆细粒棘球蚴中国大陆株诊断抗原P-29基因序列，可做为包虫病重组抗原的候选基因。

Cloning and Sequence Analysis of the Diagnostic Antigen P-29 Gene from Echinococcus Granulosus of China

Objective To obtain and analyze sequence of diagnostic antigen P-29 gene of Echinococcus granulosus. Methods Total RNA was extracted from protoscoles of cysts from human origin. The specific primers were designed according to published nucleotide sequence in the GenBank databases. The diagnostic antigen P-29 gene of Echinococcus granulosus was amplified by RT-PCR and cloned into pGEM-T vector for sequence analysis.Results A cDNA sequence with an open reading frame of 717bp had been amplified successfully by RT- PCR. Compared with DNA and amino acid sequence deduced from cDNA with the published diagnostic antigen P-29, gene sequence of Echinococus granulosus in the GenBank revealed 100% homology. Conclusion The diagnostic antigen P-29 gene sequence of Echinococcus granulosus was cloned successfully and it could be used as the candidate gene of recombinant antigen of hydatid disease.