恙虫病东方体Karp株Sta56基因的克隆及序列比较的研究

Cloning and Sequencing of the Sta56 Gene from Orientia tsutsugamushi Karp Strain

目的扩增恙虫病东方体Karp株主要表膜抗原Sta56的开放读码框(ORF)全长,并进行序列比较研究。方法小鼠接种传代和体外细胞扩增分离恙虫病东方体Karp株,用PCR方法扩增恙虫病东方体Karp株Sta56基因,并采用TA克隆技术构建测序载体,对测序结果进行同源性分析比较。结果 扩增出Sta56的ORF全长,并TA克隆到测序载体pMD18-T vector,测序结果与Karp标准株的同源性为99.4％。结论 Sta56全长ORF克隆扩增成功,为进一步构建表达载体,进行Sta56蛋白表达研究提供基础。

Objective To clone the Sta56 gene from Orientia tsutsugamushi Karp strain and analysze the DNA sequence. Method The Sta56 gene was PCR amplified, and cloned to T vector pMDIS by using TA clone technique. The DNA sequence of the gene was compared with that of the standard strain. Result The Sta56 gene was successfully amplified by PCR and the sequence showed 99.4% identity to the Standard Stain. Conclusion The successful cloning of Sta56 gene lays the foundation to further construction of expression vector and the research of expressed Sta56 protein.