›› 2007, Vol. 34 ›› Issue (9): 61-63.

• 生物技术 • 上一篇    下一篇

水貂FSH-β基因序列的克隆与测序

李玉梅, 白秀娟   

  1. 东北农业大学,哈尔滨 150030
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2007-09-20 发布日期:2007-09-20

Cloning and Sequencing on the Gene Sequence of Mink FSH-β Hormone

LI Yumei,BAI Xiujuan   

  1. Northeast Agricultural University,Harbin 150030,China 
  • Received:1900-01-01 Revised:1900-01-01 Online:2007-09-20 Published:2007-09-20

摘要: 从水貂腿部肌肉中分离提取总DNA,经PCR扩增出水貂FSH-β基因DNA序列。PCR产物经凝胶回收纯化,连接到pMD-18T载体上,然后转化到感受态细胞JM109中。在含X-gal和IPTG的LB(Amp+)平板上筛选阳性克隆菌落,提取质粒作限制性酶切及PCR鉴定后,对目的片段进行测序。结果表明, PCR 扩增出的DNA片段碱基数为500 bp,通过Blast比对,分析了其与人、牛、猪、绵羊、鼠促卵泡激素核苷酸序列的同源性。

关键词: 水貂; FSHβ基因; 克隆; 序列分析

Abstract: The author separates and extracts the overall DNA from leg muscles of mink, then gets the DNA sequence of mink FSH-β gene after PCR amplification. The product of PCR amplification is then conjunction onto pMD-18T carrier after gelatum reclaim, then transformed into competence cell JM109. bolting masculine clone colony on LB(Amp+) medium which contains X-gal and IPTG. Then after plasmid extraction, restriction enzyme cut and PCR accredit, the target segment is sequencing. As a result of the test, the amount of basic group of DNA segment after PCR amplification is 500bp, by contrast with Blast, the author analyzes the home ology of follicle-stimulating hormone ribonucleotide sequence between mink and human being, ox, pig, sheep, mouse.

Key words: mink; FSH-β gene; clone; sequence analysis

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