期刊论文

中文

中国兽医科学

1673-4696

2010

40

2

158-163

10.16656/j.issn.1673-4696.2010.02.011

湖南省科技重大专项（2007FJ1003） 湖南农业大学***基金项目（04YJ03）

本校为第一机构

根据GenBank中登录的副猪嗜血杆菌(HPS)D15基因鸟枪序列(登录号:NZ_ABKM01000005)设计1对特异性引物,以HPS 5型SP毒株DNA为模板,通过PCR方法扩增了D15基因.将其进行T-A克隆,构建pMD18D15质粒并进行序列测定和分析.结果表明,D15基因含有一个2 418 bp的开放阅读框,编码805个氨基酸,与D15基因参考序列的同源性为99.9%.该HPS D15基因的推导氨基酸序列具有典型Omp85蛋白家族的拓扑结构特征.以pMD18D15质粒为模板,用PCR方法扩增HPS D15基因,并将其克隆到pET-28a(+)中,构建pETD15原核表达质粒,将其转化大肠杆菌Rosetta(DE3)后,用IPTG诱导重组菌,并用SDS-PAGE和Western-blot检测诱导物.结果表明,pETD15重...

The full-length D15 gene was amplified from Haemophilus parasuis（HPS） serotype 5 SP strain by PCR with a pair of primers based on the whole genome shotgun sequence（Accession No.：NZ_ABKM01000005） in GenBank.Sequencing showed that the complete open reading frame（ORF） with an ATG initiation codon comprised 2 418 nucleotides and encoded a protein of 805 amino acids.It shared 99.9% identity with the reference sequence.The deduced amino acid from HPS D15 gene had the typical topology structure of Omp85 protein family.The D15 gene was successfu...