摘要
目的分析和预测牛带绦虫成虫精氨酰-tRNA转移酶（arginyl-tRNA-protein transferase,ATE）基因及其编码蛋白的结构域特性,并进行原核表达。方法利用在线生物信息学网站及工具对牛带绦虫精氨酰-tRNA转移酶基因的功能进行预测并将其编码区序列克隆到原核表达载体pET-28 a（＋）上,测序鉴定重组质粒。结果该基因全长1 476 bp,编码区为141~1 617 bp,编码491个氨基酸;该基因为全长基因,无跨膜区,具有多个磷酸化位点,蛋白的理化性质稳定,理论分子量为56 436.3Da;没有质体、线粒体定位序列;十二烷基-聚丙烯酰胺凝胶电泳（SDS-PAGE）结果表明,目的基因在大肠埃希菌BL21~DE3中表达成功。结论筛选出牛带绦虫成虫精氨酰-tRNA转移酶基因,成功构建重组原核表达质粒。
Objective To predict structural characteristics of arginyl-tRNA-protein transferase in Teania.saginata by bioinformatics and to detect its prokaryotic expression.Methods By online analysis with bioinformatics websites and software package,the function of arginyl-tRNA protein tranferase gene was predicted and the gene was inserted into the prokaryotic expression vectors pET-28a（＋） and amplified with PCR and its sequence was determined.Results A novel cDNA sequence encoding arginyl-tRNA-protein transferase with a molecular weight of 56436.3 was identified.The cDNA is 1476bp and codes 491 amino acids.PCR,double enzyme digestion and DNA sequencing indicated that pET-28a（＋） and arginyl-tRNA-protein transferase recombinant plasmid were successfully constructed.Sodium dodecyl sulfate polyacrylamide gel electrophoresis（SDS-PAGE） results showed that the gene expressed in Escherichia coli BL21/DE3.Conclusion A novel gene of Teania.saginata is successfully identified and expressed by prokaryotic vectors.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2011年第1期73-74,共2页
Chinese Journal of Public Health
基金
国家自然科学基金(30760227)
贵州省农业科技攻关〔黔科合NY字(2009-3074)〕
贵州省科技攻关(2009-3101)
贵州省省长基金〔黔省专合字(2009)82号〕
贵阳市科技局社发攻关项目(2009-3005)
