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波尔山羊生长激素(GH)基因克隆及序列分析
刘长国1, 罗 军2, 郑新民3
1.中国农科院 家禽研究所;2.西北农林科技大学 动物科技学院;3.湖北省农科院 生物工程中心
摘要:
比较生长激素(GH)基因在不同物种间的相似性.设计引物,用PCR方法从波尔山羊基因组中扩增出一条长约2.0 kb的DNA片段,然后用PMD-18T载体在感受态DH5α株大肠杆菌中获得GH基因克隆子.DNA测序结果表明,波尔山羊GH基因DNA序列全长1 951 bp(不含引物序列),含完整的5个外显子编码区.应用Clustal W软件进行同源性分析,结果表明,波尔山羊GH基因与Tokara山羊、澳洲美利奴羊和黄牛的GH基因的相似度分别为98.5%,97.0%和95.4%;内元比外元的变异程度高,在5个外元中,第一外元的保守性最高,其他区域的变异程度随物种亲缘关系的增大而升高,在分子水平上符合物种进化原理。
关键词:  波尔山羊  GH基因  克隆  序列测定  同源性分析
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基金项目:1999年度国家教委优秀青年教师基金资助项目(1511)
Cloning and sequence analysis of the Boer goat growth hormone gene
Abstract:
Based on the similarity of growth hormone gene in different species,two primers were designed and used to amplify an approximately 2.0 kb DNA fragment by PCR technique from genomic DNA sample of Boer goat.The PCR products were ligated into the PMD-18T vector,and then transformed into competent cells of E.coli DH.The DNA sequencing result showed that the GH gene of Boer goat is 1 951 bp (without primer sequence) in length and contains five complete exons.The homology analysis of the GH gene of sequences by Clustal W software indicated that the nucleotide sequence homologies of GH gene of Boer goats were 98.5% with Tokara goats,97.0% with Merino sheep,and 95.4% with bovine GH genes;variability of introns was greater than exons,the variation of other regions increased with the increasing genetic relationships of species,which is generally in agreement with the molecular evolution of species.
Key words:  Boer goat  GH gene  cloning  sequencing  homology analysis