安徽三黄鸡β-防御素Gal-6cDNA基因片段的克隆与序列分析  

Cloning and Sequence Analysis of β-defensin Gal-6gene of Anhui three-Yellow Chicken

作　　者：江龙海[1] 高恒[1] 彭开松[1] 祁克宗[1] 

机构地区：[1]安徽农业大学动物科技学院,合肥230036

出　　处：《家禽科学》2007年第10期12-15,共4页Poultry Science

基　　金：国家"863"计划(2006AA10Z320);安徽省"十五"科技重大攻关项目(0400303041);安徽省优秀青年基金(04041042)

摘　　要：根据基因库中鸡β-防御素Gal-6cDNA基因序列设计2对引物,应用反转录-聚合酶链式反应(RT-PCR)技术,从安徽三黄鸡白细胞RNA中扩增了β-防御素Gal-6cDNA基因片段。得到了大小为250bp片段。将扩增产物纯化并克隆到PGEM-T-Easy载体上,转化感受态细胞DH-5α,经筛选获得重组质粒并测序,用P3/P4引物从重组质粒中扩增出成熟肽,大小约150个bp。测序后Blast分析表明,安徽三黄鸡序列与基因库上已发表的序列相差一个碱基,Gal-6多肽共有67个氨基酸组成,分别是20个氨基酸残基的信号肽、5个氨基酸的原片段、42个氨基酸的成熟肽。According to the gene sequence of Gal-6 of chicken in Genebank,two pairs of primers were designed.Then the chicken Gal-6 cDNA fragment was amplified by reverse transcription-polymerase chain reaction （RT-PCR） from the leukocyte of three yellow chicken in Anhui.Results showed that the specific bands were found in 250bp.After gel purification,the RT-PCR products were inserted into pGEM-T-easy cloing vectors.Recombinant pGEM-T Gal-6 vector was transduced into cell DH-5α.Clones containing the DNA fragment of interest were screened,then the recombinant plasmid was sequenced.The mature peptide sequence,150bp,was amplified by the primers of P3/P4 from the recombinant plasmid.The result showed that only basic group mutated in three yellow chicken by the analysis of BLAST,the cDNA fragment code 67 amino acid residues ,comprise of signal peptide with amino acid residues,propriece peptide with 5 amino acid residues and mature oeotide with 42 amino acid residues.

关 键 词：Β-防御素 鸡 Gal-6 克隆 

分 类 号：S852.4[农业科学—基础兽医学]