摘要
目的对人禽流感病毒A/China/GD01/06（H5N1）HA基因进行克隆和表达,为进一步研究GD01/06HA的致病与免疫机制提供有效制剂。方法将H5N1全长HA基因克隆入原核表达载体pET-32a（＋）,采用异丙基-β-D-硫代半乳糖苷（IPTG）诱导表达,并对表达的重组蛋白进行鉴定。结果经检验重组质粒p32-HA构建成功,表达蛋白分子量介于66.2~94.0kd。经鉴定所表达的特异性条带为HA蛋白片段。结论HA基因在大肠杆菌中表达并获得特异性重组蛋白,为其生物学、免疫学功能的研究提供了基础。
Objective To clone the entire HA gene from HSN1 human avian influenza virus A/ China/GD01/06 and express the recombinant HA protein which was used for the further research on its pathogenic and immunologic function. Methods A full-length HA gene of A/China/GD01/06 was am- plified fiom the plasmid pMD19-T-HA which had been constructed and subcloned into the expression vector pET-32a （＋） and identified by digestion and sequencing. The recombinant plasmid p32-HA was transformed into E. coli BL21 （DE3） for expression of protein induced with IFFG. The recombinant pro- tein was identified with SDS-PAGE and western blot. Results Digestion and sequencing results of re- combinant expression plasmid p32-HA showed that prokaryotic expression vector p32-HA was success- fully constructed. The results of expression of p32-HA （BL21） displayed that the size of induced protein was between 66.2 kd and 94.0 kd. Identified results with Western blot showed that the recombinant specificity HA protein was successfully expressed. Conclusion The recombinant specificity HA protein from HSN1 human AIV is successfully obtained and can be used in the further research of its biological and immunologic function.
出处
《苏州大学学报（医学版）》
CAS
北大核心
2009年第4期604-606,624,共4页
Suzhou University Journal of Medical Science
基金
广东省科技计划项目(2005A20901005)
