农业科学与技术（英文版）
農業科學與技術（英文版）
농업과학여기술（영문판）
AGRICULTURAL SCIENCE & TECHNOLOGY
2011年
2期
195-197
,共3页
叶飞%张培君%田德雨%孙慧玲%王宏俊%龚玉梅%贺云霞
葉飛%張培君%田德雨%孫慧玲%王宏俊%龔玉梅%賀雲霞
협비%장배군%전덕우%손혜령%왕굉준%공옥매%하운하
副猪嗜血杆菌%pilA%克隆%表达
副豬嗜血桿菌%pilA%剋隆%錶達
부저기혈간균%pilA%극륭%표체
Haemophilus parasuis%pilA gene%Cloning%Expression
[目的]克隆和表达副猪嗜血杆菌外膜蛋白pilA基因.[方法]对已发表的HPS的pilA序列进行序列分析,合成引物,并以HPS血清5型基因组为模板,通过PCR扩增HPS的pilA编码基因,获得目的基因片段;构建重组表达质粒,经IPTG诱导表达至大肠杆菌BI21(DE3)中,进行SDS-PAGE与Western blot检测.[结果]表达的重组蛋白分子质量与预期的43 kD一致.[结论]为制备亚单位疫苗和诊断试剂奠定了基础.
[目的]剋隆和錶達副豬嗜血桿菌外膜蛋白pilA基因.[方法]對已髮錶的HPS的pilA序列進行序列分析,閤成引物,併以HPS血清5型基因組為模闆,通過PCR擴增HPS的pilA編碼基因,穫得目的基因片段;構建重組錶達質粒,經IPTG誘導錶達至大腸桿菌BI21(DE3)中,進行SDS-PAGE與Western blot檢測.[結果]錶達的重組蛋白分子質量與預期的43 kD一緻.[結論]為製備亞單位疫苗和診斷試劑奠定瞭基礎.
[목적]극륭화표체부저기혈간균외막단백pilA기인.[방법]대이발표적HPS적pilA서렬진행서렬분석,합성인물,병이HPS혈청5형기인조위모판,통과PCR확증HPS적pilA편마기인,획득목적기인편단;구건중조표체질립,경IPTG유도표체지대장간균BI21(DE3)중,진행SDS-PAGE여Western blot검측.[결과]표체적중조단백분자질량여예기적43 kD일치.[결론]위제비아단위역묘화진단시제전정료기출.
[Objective] The aim was to clone and express the pilA gene of outer membrane protein of Haemophilus parasuis. [Method] The published pilA gene sequence of HPS was analyzed for primer synthesis, and the genome of serotype 5-type of HPS was used as template for PCR amplification of the pilA gene of HPS; the recombinant expression plasmid was constructed and transformed into E. coli BL21 (DE3) after induced by IPTG. SDS-PAGE and Western blot analysis were then carried out. [Result] The molecular weight of expressed protein was consistent with the expected (43 kD). [Conclusion] The results provided a foundation for the preparation of subunit vaccine and diagnostic reagents.
