| 摘要: |
| 利用反转录 -聚合酶链反应 (RT-PCR) ,扩增克隆传染性法氏囊病病毒超强毒 (vv IBDV)HZ株VP2基因 ,并对 VP2基因进行全序列测定、序列分析和聚类分析 ,同时将 VP2基因与真核表达载体 pcDNA3相连接。结果克隆到 VP2基因全序列长 1 356bp,分析表明 HZ株与欧洲超强毒株 UK661高度相似 ,同时将 VP2基因正向插入 pcDNA3的 CMV启动子下游 ,得到了 VP2基因的真核表达质粒。为 IBDV分子流行病学和基因工程疫苗的研究奠定了物质基础。 |
| 关键词: IBDV,超强毒株,VP2,克隆,序列分析,真核表达质粒 |
| DOI： |
| 分类号:S852.65 9.4 |
| 基金项目:杨凌农业高新技术示范区科研基金资助项目(99KG9) |
|
| Cloning and sequence analysis of the VP2 gene in HZ strain of very virulent infectious burial disease virus and construction of its expression plasmid |
|
XU Xing gang 1 LI Jian qiang 1 WANG Xiao mei 2 TONG Guang zhi 2 LIANG Rong 1 ZHANG Yao xiang 1 XIN Fu shan 1
|
| Abstract: |
| According to the published sequence of IBDV strain 52/70, a pair of
primers that could amplify the cDNA of protective antigen VP2 gene was designed and
synthesized.By RT-PCR, a single DNA fragmentof about 1.5kb was obtained from HZ
strain of IBDV. Then the VP2 cDNA was cloned into PUC119 at SmaI site. The
nucleotide sequence of the expected VP2 gene was determined by Sanger's DNA
sequencing method, and then the amino acid sequence was deduced.Both the nucleotide
sequence and amino acid sequence were compared with five published sequence of VP2
gene of IBDV strain. It was shown that HZ strain was mostly closely related to the very
virulent strain UK661 but different from other serotype I strains.The IBDV VP2 gene
was recovered from plasmid HZ-PUC119 by digestion with XbaI. Then the VP2 gene
was subcloned into the downstream of CMV promoter of the same treated expression
vector pcDNA3.The recombinant plasmid HZ-pcDNA3 containing IBDV VP2 gene was
identified by restriction digestions and PCR amplification.The expression plasmid can be
a potentially valuable gene vaccine against IBDV. |
| Key words: infectious bursal disease virus,vvIBDV,VP2,cloning,sequence analysis,expression plasmid |
