血清Ⅰ型鸭源副粘病毒HN蛋白基因的遗传变异及原核表达  

Genetic variation and prokaryotic expression of HN gene of duck paramyxovirus

作　　者：刘艳华[1] 丁壮[1] 常爽[1] 毕玉海[1] 徐明[1] 

机构地区：[1]吉林大学畜牧兽医学院

出　　处：《中国兽医学报》2007年第3期315-318,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30571375)

摘　　要：参考GenBank发表的鹅副粘病毒（GPMV）SF02株基因组核苷酸序列，设计并合成了1对特异性引物，采用RT—PCR技术扩增鸭源血清I型禽副粘病毒DP1／0z株的HN基因，并进行遗传变异及原核表达研究。结果显示，HN基因共1716bp，编码571个氨基酸，存在6个潜在的糖基化位点和13个完全保守的半胱氨酸（C）残基。与GenBank中收录的鹕源副粘病毒HN基因核苷酸和氨基酸的同源性分别为81．3％～98．3％和87．2％～98．4％，系统进化树分析表明DP／02株与鹅源副粘病毒处于同一进化分支。经SDS—PAGE、Western—blot分析，原核表达的HN蛋白相对分子质量在63000左右，1mol／L（终浓度）IPTG时间梯度诱导，3h时HN蛋白表达量最高，占整个菌体蛋白含量的30％，且能与鼠抗鸭源血清I型禽副粘病毒阳性血清反应。According to nucleotide sequence of goose paramyxovirus （GPMV）SF02 strain,a pairs of primers was designed and synthesized. The complete HN gene of a domestic isolate DP/02 strain of duck paramyxovirus was amplified by RT-PCR, cloned into pGEMR-T vector sequenced, then explored the genetic variation and prokaryotic expression of the HN gene of DP1/02 strain. The result showed that the HN gene was 1 716 bp long and included a complete open reading frame （ORF）encoding a protein of 571 amino, containing six potential glycosylation sites and thirteen conservative cysteine residues. Compared HN gene of waterfowl paramyxovirus type 1 strains,the homology of the nucleotide and the deduced amino acids are between 81.3%-98.3% and 87.2%- 98.4% . Phylogenetic tree of HN nucleotide analysis showed that DP/02 strain has a high evolution level as goose paramyxovirus. The results of SDS-PAGE and Western-blot showed that he molecular weight of the expressed HN protein is about 63 000 and the protein amounted to 30% of the total protein of E.coli BL21（DE3）after induced by IPTG（1 mmol/L）at 37 C for 3 hours.

关 键 词：鸭 禽Ⅰ型副粘病毒 HN基因 遗传变异 原核表达 

分 类 号：Q78[生物学—分子生物学] S852.65[农业科学—基础兽医学]