猪细小病毒VP2蛋白在干酪乳杆菌表面的表达

The Surface Display of Porcine Parvovirus VP2 Protein in Lactobacillus casei

摘要:

将编码猪细小病毒主要免疫保护性抗原VP2基因插入干酪乳杆菌细胞表面表达载体pPG中，构建了重组表达载体pPG-VP2，将其电转化干酪乳杆菌Lactobacillus casei 393，获得了表达猪细小病毒VP2蛋白的重组干酪乳杆菌系统，经2%乳糖在MRS培养基中的诱导表达，SDS-PAGE检测表明，有约74kD蛋白得到了表达，表达蛋白的大小与理论值相符。Western-blot结果分析表明，表达的蛋白可被鼠源PPV抗血清所识别，间接免疫荧光实验结果表明，所表达的蛋白能够在干酪乳杆菌菌体表面检测到。

Abstract:

Lactobacillus casei 393 was selected as a bacterial carrier for the expression of Porcine Parvovirus（PPV）protective antigen VP2 protein. The gene encoding PPV VP2 protein was cloned into the Lactobacillus casei surface expression vector pPG, and then the constructed recombinant vector pPG-VP2 was electrotransformed into Lactobacillus casei 393 generating the recombinant system pPG-VP2/L.casei393 expressing PPV VP2 protein. The recombinant strain was induced by 2% Lactose in MRS and about 74kD protein was detected with SDS-PAGE. The result of Western-blot indicated that the expressed protein possessed the antigenic specificity which could be recognized by mouse anti-PPV serum. The indirect immunofluorescent test showed that the expressed protein was secreted on the cell surface Lactobacillus casei.