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五指山小型猪生长激素释放激素受体基因的cDNA克隆及序列分析
引用本文:张艳,郑心力,王峰,孙瑞萍,谭树义.五指山小型猪生长激素释放激素受体基因的cDNA克隆及序列分析[J].农业科学与技术,2009,10(4):87-90.
作者姓名:张艳  郑心力  王峰  孙瑞萍  谭树义
作者单位:海南省农业科学院畜牧兽医研究所,海南海口,571100 
基金项目:Natural Science Foundation of Hainan Province (30515).海南省自然科学基金项目 
摘    要:1材料与方法 1.1材料随机采集五指山猪品种耳组织样共30份,将其迅速放入液氮带回实验室,-70℃保存备用。 1.2试剂RT—PCRKit、PCR产物纯化回收试剂盒、重组质粒抽提试剂盒,均购自天根生物工程有限公司;SalⅠ、BamHⅠ、pMD18-TVector试剂盒、Ecoli DH5α均购自TaKaRa公司。

关 键 词:生长激素释放激素  cDNA克隆  五指山  序列分析  受体基因  小型猪  试剂盒  产物纯化

Cloning and Sequence Analysis on cDNA of Growth Hormone Releasing Hormone Receptor Gene from Wuzhishan Miniature Pig
ZHANG Yan,ZHENG Xin-li,WANG Feng,SUN Rui-ping,TAN Shu-yi.Cloning and Sequence Analysis on cDNA of Growth Hormone Releasing Hormone Receptor Gene from Wuzhishan Miniature Pig[J].Agricultural Science & Technology,2009,10(4):87-90.
Authors:ZHANG Yan  ZHENG Xin-li  WANG Feng  SUN Rui-ping  TAN Shu-yi
Institution:(Institute of Animal Science, Hainan Academy of Agricultural Sciences, Haikou 571100)
Abstract:Objective] cDNA of growth hormone releasing hormone (GHRH) receptor gene from Wuzhishan miniature pig was cloned and its sequence was also analyzed. Method] Using genomic DNA extracted from porcine ear tissues of Wuzhishan miniature pig as the template, three pairs of primers were designed by the reported cDNA sequence of porcine GHRH, and cDNA was also amplified by RT-PCR. After being recovered and purified, PCR products were ligated to pMD18-T and then transformed into Escherichia coli DH5a. The transformation products were analyzed by PCR and double enzyme digestion to screen positive clones, and the positive clones were sequenced after identification in LB liquid medium. Result] cDNA of Wuzhishan miniature pig GHRH receptor gene was obtained successfully, and its length was 1 577 bp coding 423 amino acids. BLAST analysis showed that there were only 23 nuoleotides in difference between this fragment and pomine GHRH receptor gene, and its homology was 98%. However, both GHRH receptor genes were constituted by 423 amino acids with the sequence homology of 96%. Conclusion] This study provides theoretical basis for further studies on the dwarf mechanism of Wuzhishan miniature pig.
Keywords:Wuzhishan miniature pig  Growth hormone releasing hormone receptor  cDNA clone  Analysis
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