副结核分枝杆菌SOD基因的克隆与序列分析

Cloning and Sequence Analysis of Mycobacterium paratuberculosis SOD Gene

以副结核分枝杆菌C-2染色体DNA为模板,以SOD基因特异性引物进行PCR扩增,获得约620 bp的DNA片段。将PCR产物克隆至pGEM-T Vector中,通过α-互补法筛选和质粒酶切及序列分析鉴定,成功构建出重组质粒pGEM-T-SOD,为进一步研究SOD基因及其表达产物的免疫生化特性奠定基础。

The genomic DNA was extracted from Mycobacterium paratuberculosis C2,the mature secreted protein SOD gene was amplified with a pair of specific primers through using polymerase chain reaction（PCR）.The PCR product was approximately 620 bp DNA segment.The PCR product was cloned into pGEM-T vector,then by using α-complementation test,restrictional enzyme assay and recombinant plasmid sequence analysis,Recombinant plasmid pGEM-T-SOD was successfully constructed.These results could serve as a basis for further studies on the usefulness of SOD gene and immunogenicity of SOD gene expression product.