摘要
用大片吸虫成虫FG0018表达序列标签(Expressed Sequence Tags,ESTs)序列对NCBI数据库进行Blastn比较,采用生物信息学方法,运用网上数据库资源,结合相关生物信息学软件将目标序列进行搜索、处理、拼接、延伸,得到的最后重叠群(contig,命名为FG0018C0N)与肝片吸虫卵壳蛋白B2序列(GenBank:M93025)相似性很高,用DNASTAR、SEQMAN程序和Blast2程序分析FG0018C0N,推测其开放阅读框为930 bp,编码310个氨基酸,蛋白质的特点及跨膜区预测其编码蛋白可能属于核蛋白,疏水性分析表明其编码一疏水蛋白。FG0018C0N基因与肝片吸虫卵壳蛋白基因有100%的相似性,310个氨基酸与之有91%的一致性。经RT_PCR、酶切验证并cDNA测序,结果证明FG0018C0N序列可能是大片吸虫的卵壳蛋白基因。
The expressed sequence tags of FG0018 of Fasciola gigantica was subjected to Blastnsearching against the parasite dbEST and Htgs in NCBI., After searching, assemblying and extending by use of the bio-softwares, bioinformatic methods, internet database,a contig named as FG0018CON was obtained. It was high homologous with the Fasciola hepatica vitelline protein B2 mRNA, and then take use of the DNASTAR,SEQMAN and Blast2 programe to predict it＇s ORF and some functions , results show it＇s ORF is 930 bp which encodes a protein of 310 Amino Acids and predicted as a trans-membrane, hydrophobe and neuclear protein. The FG0018CON shows 100 % identity in 868bp overlap with the Fasciola hepatica vitelline protein 132 mRNA gene and the predicted polypeptide shows 91% identity in 310 aa with the Fasciola hepatica vitelline protein. Then the FG0018CON was verifed that it is a vitelline gene of Fasciola gigantica by the RT-PCR, Digestion and sequencing methods.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2005年第5期418-422,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
国家自然科学基金(30260082):广西科学基金(桂科基0448001)
关键词
生物信息学
大片吸虫
电子克隆
表达序列标签
bioinformatics
Fasciola gigantica
in sihco cloning
expressed sequence tag
