2拷贝SLT-ⅡvB在猪霍乱沙门氏菌C500crp^-/asd^-缺失株中的表达及小鼠免疫试验

Expression of 2 Copies SLT-ⅡvB Gene in C500crp^-/asd^- Mutant of Salmonella choleraesuis and Immune Test on Mice

为研制仔猪副伤寒—猪水肿病新型二联疫苗,从pMDSLT-Ⅱ中扩增去信号肽后的SLT-ⅡvB基因,将2拷贝SLT-ⅡvB与pYA3493连接。阳性质粒pYA2B电转化C500crp-/asd-缺失株,进行SDS-PAGE电泳和Western blotting,重组菌C500crp-/asd-(pYA2B)口服免疫小鼠,腹腔攻10 LD50的猪霍乱沙门氏菌强毒C78-1和2 LD50的猪水肿大肠杆菌强毒ED3株,计算攻毒保护率。结果显示,2 SLT-ⅡvB在C500crp-/asd-缺失株中获得了表达。C500crp-/asd-(pYA2B)免疫组对ED3的攻击能提供60%保护,而对照组不能提供任何保护。2组对C78-1攻击均提供100%保护,为研制仔猪副伤寒—猪水肿病二联疫苗及进一步提高免疫效果提供依据。

In order to develop novel piglet＇s Paratyphoid-ED bivalent vaccine,SLT-ⅡvB gene without signal peptide was amplified from pMDSLT-Ⅱ and subcloned into pYA3493.pYA2B was electroporated into C500crp-/asd-mutant.SDS-PAGE and Western blotting were done.Mice were i.p.immunized with C500crp-/asd-（pYA2B） and C500crp-/asd-（pYA3493） and i.p.challenged with 2 LD50 ED3 and 10 LD50 C78-1.2 SLT-ⅡvB was expressed in C500crp-/asd-mutant.The C500crp-/asd-（pYA2B） immunized group was 60% protected mice against ED3 challenging and control groups couldn＇t protect mice.All immunized groups induced 100% protected mice against C78-1 challenging.These results provided base for developing piglet＇s Paratyphoid-ED bivalent vaccine and further improving of immunity effect.