抗菌肽Protegrin-1基因的PCR扩增和克隆及序列分析  

PCR amplication, cloning and sequencing of Protegrin-1 gene of porcine antibacterial peptide.

作　　者：汪以真[1] 刘光富[1] 初晓娜[1] 韩菲菲[1] 

机构地区：[1]浙江大学饲料科学研究所,浙江杭州310029

出　　处：《浙江大学学报（农业与生命科学版）》2004年第3期336-339,共4页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：国家自然科学基金资助项目(30040036);浙江省人才培养专项基金资助项目(RC01053).

摘　　要：从杜长嘉猪的骨髓中提取基因组RNA,用反转录-聚合酶链式反应(RT-PCR)扩增抗菌肽Protegrin-1(PPG-1)基因,获得一条565bp的片断,以pGEM-TEasyvector为载体,将该基因片断克隆到大肠杆菌中.从筛选到的阳性克隆中分离出Protegrin-1基因,测定其序列,并由此推导出氨基酸序列.结果表明:1该片断长565bp,为Protegrin-1基因cDNA的部分序列,序列同源性比较表明,该基因序列与GeneBank登录的Protegrin-1基因序列具有较高的同源性,达到98.8%.2根据所测序列推导其成熟肽由18个氨基酸残基组成,其氨基酸序列与GeneBank登录的Protegrin-1的成熟肽序列完全一致.Genome RNA was extracted from bone marrow of pig (Duroc╳Landrace╳Tai-hu) and Protegrin-1 (PPG-1) mRNA was amplified using RT-PCR. A DNA fragment about 565 bp in length was obtained and the PCR product was cloned into pGEM-T vector. Protegrin-1 gene was isolated and sequenced from the positive clones screened and the corresponding amino acid sequence was deduced from its nucleotide sequence. Result of sequence analysis showed that this fragment was the partial sequence of PPG-1 gene cDNA. Its homology with PPG-1 from Genebank was up to 98.7%.The deduced sequence for the mature peptide was composed of 18 amino acid residues, which was consistent with that of porcine PPG-1 in Genebank.

关 键 词：猪 骨髓 PPG-1 RT—PCR 基因克隆 序列分析 

分 类 号：Q785[生物学—分子生物学]