摘要
目的 :对鼠疫耶尔森氏菌F1抗原在鼠伤寒沙门氏菌中的表达及其保护力进行研究。方法 :PCR、化学转化及电穿孔法。结果 :采用PCR方法 ,特异地扩增了鼠疫菌F1抗原caf操纵子的 5Kb基因片段。将PCR产物与质粒载体连接后 ,经化学方法转化大肠杆菌LE392 ,获得caf基因克隆子 ,所得克隆子能较好地表达F1抗原基因。应用电穿孔法将克隆子的重组质粒转入鼠伤寒沙门氏菌G30内获得转化子 ,其F1抗原表达水平与在大肠杆菌LE392中的表达水平相同 ,反向血凝滴度可达 1∶80以上 ,与EV株相近。用该株免疫小鼠 ,2 0d后用 141株攻击 ,可延长平均死亡时间。结论 :caf基因的克隆子在鼠伤寒沙门氏菌G30中能够有效表达 ,且对小鼠显示出保护效果。
The caf operon encoding F1 antigen of Y.pestis was amplified by PCR method, The PCR products were ligated to a vector plasmid, then transformed into Ecoli LE392. Some clones with caf operon obtained. These clones could express efficiently F1 antigen. The recombinant plasmids of those clones were extracted and transformed into S.typhimurium G30 by electroporation, some transformants were obtained, their expression level of F1 antigen was similar to those of Ecoli LE392 clones. The titer was up to 1∶80 as detected by IHA. These clones with caf operon could express efficiently in S. typhimurium , The clone immunized mice,the mice challenged with virulent 141 strain after 20 days. The survival was prolonged over that of negative control, our study provides pilot.
出处
《中国媒介生物学及控制杂志》
CAS
CSCD
2000年第4期293-297,共5页
Chinese Journal of Vector Biology and Control
