期刊文献+

柔嫩艾美耳球虫杨凌株TA4基因的克隆和原核表达

Cloning and prokaryotic expression of TA4 gene from E.tenella YL strain
原文传递
导出
摘要 根据国外报道的序列设计1对引物,以纯化的柔嫩艾美耳球虫(E.tenella)杨凌(YL)株孢子化卵囊的总RNA为模板,用RT-PCR方法扩增出E.tenella YL株的TA4基因。序列分析表明,该序列全长741bp,开放阅读框(ORF)为651bp,共编码217个氨基酸,与E.tenella HT株、E.tenella BJ株和E.tenella ZJ株的TA4基因ORF序列同源性分别为99.8%、99.8%和99.27%。由此确定所获得的目的基因为E.tenella YL株TA4基因。利用生物信息学和分子生物学软件对TA4基因编码的蛋白进行结构预测,结果表明,该蛋白为一结构松散的球状蛋白。将TA4基因克隆到表达载体pET-32a中,构建重组质粒pET-TA4并在大肠杆菌BL21中进行表达,表达产物的SDS-PAGE分析结果表明,成功地表达了分子质量为41ku的融合蛋白。 A pair of primers was designed according to TA4 gene sequence of Eimeria tenella HT strain available in GenBank.Then a TA4 gene was amplified using total RNA from the purified E.tenella YL strain sporulated oocysts as template by RT-PCR.Sequence analysis showed that ORF of TA4 gene was consisted of 651 nucleotides and encoded 217 amino acids.Compared with E.tenella HT strain,E.tenella BJ and E.tenella ZJ strain,TA4 gene from E.tenella YL strain shared 99.8%,99.8% and 99.27% homology,respectively.The protein structure prediction showed that the TA4 protein was a loose globe protein.The recombinant expression plasmid pET-TA4 was constructed and expressed in BL21 strain of Escherichia coli.SDS-PAGE analysis showed that the fusion protein of 41ku in molecular weight was successfully expressed.
出处 《中国兽医科学》 CAS CSCD 北大核心 2010年第5期481-485,共5页 Chinese Veterinary Science
基金 陕西省农业攻关项目(2008K02-06) 985工程科技创新平台项目(Z101020001)
关键词 柔嫩艾美耳球虫 TA4基因 克隆 表达 结构预测 Eimeria tenella TA4 gene cloning expression structure prediction
  • 相关文献

参考文献12

二级参考文献75

  • 1吴绍强,蒋金书,刘群,朱引洁.柔嫩艾美耳球虫BJ株TA4基因的表达及电泳分析[J].中国农业科学,2004,37(8):1217-1221. 被引量:3
  • 2田广孚,田增义.柔嫩艾美尔球虫子孢子的提纯[J].中国兽医杂志,1989,15(4):27-28. 被引量:11
  • 3简永利,蔡建平,于三科,覃宗华,林青,叶秀华,陈闻,党海斌.柔嫩艾美耳球虫表面抗原SAG2基因的克隆与表达[J].畜牧与兽医,2006,38(6):10-13. 被引量:4
  • 4SHIRLEY M W,SMITH A L,BLAKE D P,et al.Changes in successful control of the avian coccidian[J].Vaccine,2007,25(30):5540-5547.
  • 5LILLEHOJ H S,CHOI K D,JENKINS M C,et al.A recombinant Eimeria protein inducing interferon-γ production:comparison of different gene expression systems and immunization strategies for vaccination against coccidiosis[J].Avian Dis,2000,44(2):379-389.
  • 6JOHNSON J,REID W M.Anticoccidial drugs;lesion scoring techniques in battery and floor-pen experiments with chickens[J].Exp Parasitol,1970,28(1):30-36.
  • 7LILLEHOJ H S,DING X,DALLOUL R A,et al.Embryo vaccination against Eimeria tenella and E.acervulina infections using recombinant proteins and cytokine adjuvant[J].J Parasitol,2005,91(3):666-673.
  • 8KLOTZ C,GEGRE F,LUCIUS R,et al.Identification of Eimeria tenella genes encoding for secretory proteins and evaluation of candidates by DNA immunization studies in chickens[J].Vaccine,2007,25(36):6625-6634.
  • 9XU Q,SONG X,XU L,et al.Vaccination of chickens with a chimeric DNA vaccine encoding Eimeria tenella TA4 and chicken IL-2 induces protective immunity against coccidiosis[J].Vet Parasitol,2008,156(3/4):319-323.
  • 10DU A,WANG S.Efficacy of a DNA vaccine delivered in attenuated Salmonella typhimurium against Eimeria tenella infection in chickens[J].Int J Parasitol,2005,35(7):777-785.

共引文献92

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部