期刊论文

中文

中国兽医科学

1673-4696

2010

40

8

822-826

10.16656/j.issn.1673-4696.2010.08.009

(2006BAD06A00):国家科技支撑计划项目 (K0902007-21):长沙市科技计划重大专项

本校为第一机构

为研究副猪嗜血杆菌(HPS)基因工程疫苗,以HPS 5型毒株为模板,通过PCR扩增获得副猪嗜血杆菌神经氨酸酶(Neu)全基因,将其克隆至pMD18-T载体并对其进行测定和分析.结果表明,Neu全基因含一个2187 bp的开放阅读框,编码一含729个氨基酸的蛋白,与Neu基因参考序列(登录号:NZABKM01000006)的同源性为99.0%.以pMD-Neu基因为模板,重新设计引物,扩增得到HPS Neu蛋白的部分编码区,将其克隆至原核表达载体pET-28a(+)中,构建得到重组原核表达质粒pET-Neu.将pETNeu在大肠杆菌Rosetta(DE3)中进行优化表达,经SDS-PAGE分析,重组蛋白Neu主要以包涵体形式表达,分子质量为52 ku;Western-blot分析表明,重组蛋白Neu能与阳...

The full-length Neu 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 amplified Neu gene was successfully cloned into pMD18-T vector and the complete open reading frame with an ATG initiation codon comprised 2187 nucleotides and encoded a protein of 805 amino acids.It shared 99.0% identity with that of the reference sequence.The HPS Neu gene fragment was amplified by PCR from pMD...