›› 2009, Vol. 36 ›› Issue (9): 59-62.

• 生物技术 • 上一篇    下一篇

扩展莫尼茨绦虫引发酶蛋白基因的克隆和cDNA序列分析

吕廷德1,2,赵文娟2,朱建军2,王新华2,薄新文2   

  1. (1.石河子大学, 石河子 832000; 2.新疆农垦科学院兵团绵羊繁育生物技术重点实验室, 石河子 832000)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-09-20 发布日期:2009-09-20
  • 通讯作者: 薄新文

Cloning and Sequence Analysis of the cDNA Encoding Primase in Moniezia expansa

LV Ting-de1,2, ZHAO Wen-juan2, ZHU Jian-jun2, WANG Xin-hua2, BO Xin-wen2   

  1. (1.Shihezi University, Shihezi 832000, China; 2.The Breed and Biotechnology Key Laboratory of Sheep in Xinjiang Bingtuan, Shihezi 832000, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-09-20 Published:2009-09-20
  • Contact: BO Xin-wen

摘要: 分离和鉴定扩展莫尼茨绦虫(Moniezia expansa)新基因,为进一步研究该基因的功能奠定基础。构建扩展莫尼茨绦虫成虫cDNA文库,随机挑取重组阳性克隆进行测序,对部分序列进行引物步移法测序,获取其全长cDNA序列;采用生物信息学等分析技术对该cDNA序列进行开放阅读框(ORF)的寻找、编码氨基酸的推导、核苷酸和氨基酸同源性比较及蛋白质二级结构的初步预测。获得了1个扩展莫尼茨绦虫新基因——引发酶蛋白,全长1269 bp,编码422个氨基酸,属于AE_Prim_S家族。编码蛋白的理论分子质量为47.1598 ku,等电点为4.83。获得了扩展莫尼茨绦虫反应结合蛋白的全长cDNA序列,为该基因功能的试验性鉴定工作奠定基础。

关键词: 扩展莫尼茨绦虫; 引发酶; 序列分析; 表达序列标签

Abstract: To clone and identify novel genes from an adult Moniezia expansa (M.expansa) cDNA library, and provide a foundation for further research, a cDNA library was constructed from M.expansa adult stage. Clones were selected randomly from the cDNA library and were sequenced by using the method of expression sequence tags (ESTs). Novel genes were acquired by primer-walking. The cDNA sequence encoding M.expansa primase protein was analyzed, including searching the ORF, translating the nucleotide to protein sequence, similarity searches and secondary structure predication with bioinformatics analysis. Primase genes, 1269 bp and coding for 422 amino acids, was cloned and sequenced, then the sequence was submitted to GenBank and got an accession number,GH291475. The theoretical pI was 4.83 and molecular weight was 47.1598 ku. The full-length cDNA sequence encoding M.expansa primase was obtained, which gave a basis for further functional study of this gene.

Key words: M.expansa; primase; sequence analysis; ESTs

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