摘要： 大肠杆菌不耐热性肠毒素(LT)具有免疫增强作用,是良好的免疫佐剂。为了更好的研究其免疫增强效果,本研究将LT的两个亚基LTa(R210G)和LTb克隆到pGST中,原核表达LTa(R210G)和LTb,经Western-blotting分析,表达蛋白具有免疫原性。对表达蛋白进行大量纯化后,以卵清蛋白为模式蛋白(OVA)为模式蛋白,免疫小鼠,ELISA结果表明,OVA+LTa(R210G)佐剂组小鼠的抗体滴度最高,OVA+LTb和OVA+LTa+LT组抗体滴度相似,高于OVA+氢氧化铝组,而OVA+氢氧化铝组高于无佐剂OVA组。因此,LTa(R210G)和LTb具有良好的免疫增强作用,是一种极有希望的免疫佐剂。
关键词:
大肠杆菌不耐热性肠毒素,
LTa和LTb,
原核表达,
免疫反应
Abstract: Ecoli heat -labile enterotoxin is a best immune adjuvant for its immunic enhancement.In order to studythe effect of its immunic enhancement,pGST -LTa(R210G)and pGST -LTb was constructed and expressed,the resultof western -blotting showed the recombinant protein of GST -LTa(R210G)and GST -LTb had good biological activitiesanalogy to LT positive sera from pigs.We immunized mice with Ovalbumin,using GST -LTa(R210G)and GST -LTb asimmunoadjuvant,and aluminum hydroxide adjuvant as controls.GST -LTa(R210G)showed the capability to remarkablyincrease the level of antibody against ovalbumin when comparing with those mice without any immunoadjuvant,and whichwas even higher than that of aluminum hydroxide adjuvant,LTb and LTa (R210G)plus LT adjuvant.LTb and LTa(R210G)plus LT adjuvant had the same level of antibody against Ovalbumin.Therefore,LTb and LTa (R210G)can becomea new type of immunoadjuvant in future.
Key words:
Ecoliheat-labileenterotoxin,
LTaandLTb,
Prokaryoticexpression,
Immunogenecity
中图分类号:
YUANXiu-fang, WANGYi-cheng, XULi-hua, LIUDeng, NIUDeng-yuan, WANGChao-wen. Prokaryotic Expression of Ecoli Heat -labile Enterotoxin LTa and LTb and Its Immunogenenity in Mice[J]. ACTA AGRICULTURAE BOREALI-SINICA, doi: 10.7668/hbnxb.2009.S1.062.