【摘要】 应用重组PCR的方法,直接从盐沼盐杆菌(Halobactrium salinarium)S9的基因组DNA上扩增到了编码细菌视紫红质(bacteriorhodopsin)的基因———bop基因及其启动子部分(1 196 bp),并将其克隆到表达载体pXLNovR后,在宿主菌株中得到了表达.测序结果表明,扩增得到的片段与NCBI数据库中的bop基因及启动子的序列完全相同,且其表达产物的分子量与野生型BR蛋白的分子量一致,并且在568 nm处表现出BR蛋白特征吸收峰.本方法与其他方法(如逆转录PCR,建立基因文库后从中调取等方法)相比具有操作简便,成功率高的优点.更多还原

【Abstract】 In the present study, the bop gene and the promotor were amplified directly from the genomic DNA of Halobactrium salinarium S9 by recombinant-PCR method. Then, we constructed a recombinant plasmid pXLNovR-bop and expressed bop gene in the L33 cells. The length of the sequence is1196 base pair which is the same with the sequence of bop gene in the NCBI database. The molecular weight of the BR protein expressed from the cloned bop gene is also the same with the BR protein of the Halobactrium salinarium S9 which shows a characteristic absorption peak located at 568nm.This method is easier and reliable than other methods.更多还原