共表达KnobS蛋白与LTB蛋白的重组干酪乳杆菌构建

在干酪乳杆菌中表达减蛋综合征病毒纤维蛋白KnobS,并分析其抗原性。分别将编码减蛋综合征病毒主要免疫保护性抗原纤维蛋白KnobS与大肠杆菌不耐热肠毒素B亚单位(LTB)基因插入干酪乳酸杆菌分泌表达载体pMG36e-UP/L中,成功构建重组表达载体pMG36e-UP/LTB-KnobS,获得表达融合蛋白KnobS-LTB的重组乳酸菌干酪乳杆菌表达系统;经SDS-PAGE和Western-blot检测表明,有大小约41kD的蛋白表达,具有与天然病毒蛋白一样的抗原特异性。在干酪乳杆菌中成功地表达了含减蛋综合征病毒抗原的融合蛋白KnobS-LTB,且具有较好的抗原性,为减蛋综合征病毒重组乳酸菌活菌口服疫苗的研制奠定重要的物质基础。

Co-expression of EDSV KnobS and LTB on the Surface of Lactobacillus Casei

We tested the antigenicity of egg drop syndrome virus （EDSV） KnobS gene expressed in Lactobacillus. The fusion protein gene encoding EDSV KnobS protein and LTB, was cloned into the surface expression vector pMG36e-UP/L, and then the recombinant expression vector pMG36e-UP/ LTB-KnobS was electrotransformed into Lactobacillus casei 393, generating recombinant strain pMG36e-UP/LTB-KnobS/Lactobacillus casei 393. After induced in MRS broth, an about 41kD pro- tein was detected in the recombinant Lactobacillus casei by SDS-PAGE. The result of Western blot indicated that the protein possessed the antigenic specificity same as the native virus protein. KnobS- LTB gene was expressed in Lactobacillus successfully. It had good immunogenicity and protection a- gainst porcine rotavirus. The results provide potential for the development of lactic acid bacteria oral vaccine of EDSV,which used LTB as mucosal adjuvant.