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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。
OBJECTIVE: To clone the outer membrane protein Omp38 of Burkholderia pseudomallei and to identify the antigenicity of the recombinant Omp38 (rOmp38) protein. Methods: PCR was used to amplify the Omp38 gene of Burkholderia pseudomallei. The amplified product was cloned into the expression vector pET-28a (+). After the recombinant expression vector was identified, the recombinant protein was induced to express and purify, and the antigenicity of the recombinant protein was identified by Western blotting. RESULTS AND CONCLUSION: The recombinant plasmid pET-28a / Omp38 was constructed and Omp38 gene was highly expressed. Western blotting showed that rOmp38 protein has good antigenicity.