摘要
目的获得细粒棘球蚴(E.granulosus)铁蛋白(ferritin)基因序列,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,根据国外细粒棘球蚴铁蛋白基因的已知序列设计一对引物,采用RT-PCR技术扩增出细粒棘球蚴中国大陆株铁蛋白基因,待PCR产物纯化后,将其克隆到pGEM-T载体,进行序列测定和分析。结果用RT-PCR成功扩增出细粒棘球蚴中国大陆株铁蛋白基因,测序表明该基因为562bp。同源性比较结果表明:该基因序列与已发表基因核苷酸序列相比,390bp同源性约为97.7%,推导编码氨基酸序列同源性为97.7%,此基因序列在435bp出现终止密码。结论测序的铁蛋白基因序列有完整的编码框,为进一步研究其结构和功能,以及对包虫病的免疫预防具有重要意义。
Objective To obtain and analyze the sequence of ferritin gene. Methods Total RNA was extracted from protoscolexes of cysts from human origin. The specific primers were designed according to published nucleotide sequence in the Genebank database. The ferritin gene of Echinococcus granulosus was amplified by RT- PCR and cloned into pGEM-T vector for sequencing and analyzing. Results A cDNA sequence of 562bp has been amplified successfully by RT-PCR. Comparision 390bp of the DNA and amino acid sequence deduced from cDNA with the published ferritin gene sequence of Echinococcus granulosus in the Genebank revealed identity of 97.7 %, it had a terminate code at 435bp. Conclusion The ferritin sequence gene has a completely open reading frame. E. granulosus ferritin gene gained from protoscolexes can be used as candidate antigen gene to develop vaccine and further biological functions studying.
出处
《宁夏医学院学报》
2005年第4期253-255,261,共4页
Journal of Ningxia Medical College
基金
国家自然科学基金(30260105)
关键词
细粒棘球蚴
铁蛋白
基因
克隆
序列分析
echinococcus granulosus
ferritin
gene
cloning
sequencing
