类鼻疽伯克霍尔德菌Omp38基因的克隆表达及表达产物抗原性鉴定

目的：克隆表达类鼻疽伯克霍尔德菌外膜蛋白Omp38,并对重组Omp38（rOmp38）蛋白的抗原性进行鉴定。方法：应用PCR技术对类鼻疽伯克霍尔德菌Omp38基因进行特异性扩增,将扩增产物克隆入表达载体pET-28a（＋）。重组表达载体经鉴定后,对重组蛋白进行诱导表达纯化,利用Western印迹对重组蛋白的抗原性进行鉴定。结果与结论：构建了pET-28a/Omp38重组表达载体,Omp38基因得到高效表达,Western印迹显示rOmp38蛋白具有较好的抗原性。

Cloning and expression of Omp38 gene from Burkholderia pseudomallei and its immunogenicity characterization

Objective：To clone and express the Omp38 outer membrane protein gene of Burkholderia pseudomallei and to identify the immunogenicity of the recombinant protein. Methods ： The omp38 gene of B. pseudomallei was specifically am- plified by PCR and cloned into prokaryotic expression vector pET-28a （ ＋ ）, and the recombinant plasmid was transferred into competent E. coli BL21 （DE3）. Positive clones were screened and identified by direct colony PCR, restriction enzyme digestion and sequence analysis. Then the recombinant vector was induced and expressed in E. coli BL21 （DE3）. The im- munogenicity of the recombinant protein was assessed by Western-blot. Results and Conclusion ：The recombinant expres- sion vector pET-28a/Omp38 was successfully constructed, and the recombinant Omp38 protein was highly expressed in the E. coli BI21. Western-blot analysis showed the recombinant protein exhibited a good immunogenicity.