牛带绦虫ATE基因功能预测及原核表达
Function and prokaryotic expression of arginyl-tRNA-protein transferase in Teania, saginata
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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.Abstract: Objective To predict structural characte ristics of arginyl tRNA prote in transferase in Teania.saginata by bioinformatics and to detectits prokaryotic expression.Methods By online analysis with bioinform atics websites and soft ware package, the function of arginyl tRNA protein tranferase gene was predicted and the gene was inserted into the prokary otic expression vectors pET 28a(+) and amplified with PCR and its sequence was determined.Results Anovel cDNAse quence encoding arginyl tRN A 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 RtNA prote in transferase recombinant plasmid were successfully constructed.Sodium dodecyl sulfate poly acry lamide gelelectrophoresis(SDS PAGE) results showed that the gene expressed in Escherichia coli BL 21/DE3.Conclusion Anovel gene of Teania.saginata is successfully identified and expressed by prokaryotic vectors.
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