禽巴氏杆菌株粘附蛋白基因的克隆和序列分析

doi:10.3969/j.issn.1007-2985.2015.02.015

journal6 ›› 2015, Vol. 36 ›› Issue (2): 77-81.DOI: 10.3969/j.issn.1007-2985.2015.02.015

禽巴氏杆菌株粘附蛋白基因的克隆和序列分析

段世雄，吾鲁木汗·那孜尔别克，恩特马克·布拉提白

（1.吉首大学生物资源与环境科学学院，湖南吉首 416000；2.伊犁师范学院化学与生物科学学院，新疆伊宁 835000）

出版日期:2015-03-25 发布日期:2015-04-28
通讯作者: 恩特马克·布拉提白（1961—），男，新疆伊犁昭苏县人，伊犁师范学院化学与生物科学学院教授，博士，主要从事动物病原微生物学研究.
作者简介:段世雄（1984—），男，湖南长沙人，吉首大学生物资源与环境科学学院硕士研究生，主要从事微生物分子生物学研究
基金资助:
国家自然科学基金资助项目（30972206,31360613）

Cloning and Sequencing of Adhesive Protein Genes of Avian Pasteurella Multocida

DUAN Shixiong,NAZIERBIEKE Wulumuhan,BORRATHYBAY Entomack

(1.College of Biology and Environmental Sciences,Jishou University,Jishou 416000,Hunan China;2.College of Chemistry and Biological Sciences,Yili Normal University,Yining 835000,Xinjiang China)

Online:2015-03-25 Published:2015-04-28

摘要/Abstract

摘要：用PCR从禽巴氏杆菌P-1059基因组DNA中分别扩增编码粘附蛋白OmpH，PtfA，PlpE和PM1665的基因序列，将PCR产物克隆到pMD18-T载体中，转化大肠杆菌DH5α，经菌落PCR筛选含有重组质粒DNA的重组子，并对目的基因进行测序.DNA测序结果表明，P-1059株ompH，plpE，ptfA和pm1665基因大小分别为1 032，1 008，435，348 bp，分别编码343，335，144，115个氨基酸的多肽.Blast分析结果显示，P-1059株与已报道不同血清型巴氏杆菌ompH基因的同源性为95%~97%，与ptfA基因的同源性为95%~99%，与plpE基因的同源性为94%~97%，而与pm1665基因的同源性为99%.克隆得到的ompH，ptfA，plpE和pm1665基因，为禽巴氏杆菌粘附因子及其致病机理的研究奠定基础.

关键词： 禽多杀性巴氏杆菌, 粘附蛋白, 基因克隆, 核苷酸序列同源性

Abstract: The genes encoding the outer membrane protein H (OmpH),lipoprotein E (PlpE),type 4 fimbrial subunit protein (PtfA) and fibronectin binding protein (PM1665) were amplified by PCR from genomic DNA of avian Pasteurella multocida P-1059,respectively.The PCR products were cloned into the pMD18-T vector and then sequenced.Sequence analyses showed that the open reading frames (ORFs) of the ompH,plpE,ptfA and pm1665 genes were 1 032,1 008,435 and 348 bp in length,and encoded the 343,335,144 and 115 amino acid residues.Nucleotide sequence homologies of the ompH,plpE,ptfA and pm1665 genes between the P-1059 and the previously reported different serotype strains of P. multocida were 95%~97%,95%~99%,94%~97% and 99%,respectively.In conclusion,the cloned genes encoding the adhesive proteins of avian P. multocida P-1059 will contribute to further study on pathogenesis of this organism.

Key words: Pasteurella multocida, adhesive protein, gene cloning, nucleotide sequence homology

段世雄, 吾鲁木汗·那孜尔别克, 恩特马克·布拉提白. 禽巴氏杆菌株粘附蛋白基因的克隆和序列分析[J]. journal6, 2015, 36(2): 77-81.

DUAN Shixiong,NAZIERBIEKE Wulumuhan,BORRATHYBAY Entomack. Cloning and Sequencing of Adhesive Protein Genes of Avian Pasteurella Multocida[J]. journal6, 2015, 36(2): 77-81.

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[13] 吾鲁木汗·那孜尔别克,张宇凤,龚凤娟,等.禽多杀性巴氏杆菌C48-3株ompH基因敲除突变株的构建及其生物学功能[J].微生物学报,2013,53(1):66-73.

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禽巴氏杆菌株粘附蛋白基因的克隆和序列分析

Cloning and Sequencing of Adhesive Protein Genes of Avian Pasteurella Multocida

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