›› 2008, Vol. 1 ›› Issue (2): 60-63.

• 生物技术 • 上一篇    下一篇

小尾寒羊雌激素受体基因外显子4的克隆与序列分析

贾立华1,储明星2,陈宏权1,方丽2
  

  1. 1.安徽农业大学动物科技学院,合肥 230036; 2.中国农业科学院北京畜牧兽医研究所,中国农业科学院家养动物遗传资源与种质创新重点开放实验室,北京 100094
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-02-20 发布日期:2008-02-20

Cloning and Sequence Analysis of Exon 4 of Estrogen Receptor Gene in Small Tail Han Sheep

JIA Lihua1, CHU Mingxing2, CHEN Hongquan1, FANG Li2
  

  1. 1. College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;2. The Key Laboratory of Domestic Animal Genetic Resources and Germplasm Innovation of CAAS, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100094, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-02-20 Published:2008-02-20

摘要: 根据GenBank发表的人、鸡、大鼠雌激素受体(estrogen receptor,ESR)基因外显子4的序列设计1对引物,采用PCR 技术扩增出小尾寒羊ESR基因外显子4的DNA片段,将该片段克隆到pGEM-T Easy 质粒中,重组质粒用PCR 扩增进行阳性克隆鉴定,然后测定其核苷酸序列并推导氨基酸序列,同时将测定的小尾寒羊ESR基因外显子4序列与人、牛、猪、大鼠、鸡的外显子4序列进行比较。结果表明:克隆测序所得的核苷酸和翻译后的氨基酸序列与人、牛、猪、大鼠、鸡相比,同源性分别为77.68%~97.28%和71.82%~98.18%,显示了很强的保守性;小尾寒羊的核苷酸序列与人、牛、猪、大鼠、鸡相比存在1处特有变异,氨基酸序列23~36存在高变异区。

关键词: 小尾寒羊; 雌激素受体基因; 克隆; 序列分析

Abstract: According to the human, chicken and rat sequence of exon 4 of estrogen receptor (ESR) gene, a pair of primer was designed. DNA fragment of exon 4 of Small Tail Han sheep ESR gene was amplified successfully by PCR, and was cloned into pGEM-T Easy vector. The positive clones were further identified by PCR. The nucleotide sequence was detected and the amino acid sequence of this fragment was deduced. The comparison of the fragment of exon 4 of Small Tail Han sheep ESR gene with that of human, cattle, pig, rat and chicken ESR gene showed that the homologies of the nucleotide sequence and amino acid sequence of the fragment with human, cattle, pig, rat and chicken were from 77.68% to 97.28% and from 71.82% to 98.18%, respectively, which indicated that the ESR gene was highly conserved. Nucleotide sequence existed one special variation and the peptide sequence of amino acids 23 to 36 was highly variable in Small Tail Han sheep.

Key words: Small Tail Han sheep; estrogen receptor gene; cloning; sequence analysis

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