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先以含全长SR-BIcDNA序列的重组质粒pMD18-T-rS为摸板进行PCR反应扩增SR-BI得到胞外域cDNA片段,经测序证明正确后,定向克隆到酵母双杂交表达载体,然后与pGBKT7-ApoA-I质粒共转化酵母细胞,通过报告基因及酵母交配试验确认了SR-BI的胞外域部分和ApoA-I之间的确存,观察到ApoA-I与SR-BI胞外域间的相互作用力比与全长的SR-BI间的相互作用力提高了10%。
The recombinant plasmid pMD18-T-rS containing the full-length SR-BIcDNA sequence was used as a template for PCR amplification of SR-BI to obtain the extracellular domain cDNA fragment. After sequencing proved correct, the recombinant plasmid was cloned into the yeast two-hybrid expression vector Yeast cells were co-transformed with the pGBKT7-ApoA-I plasmid and the presence of the extracellular domain between ApoA-I and SR-BI was confirmed by the reporter gene and yeast mating assay. ApoA-I was observed between the extracellular domain of SR- The interaction force increased by 10% with the full-length SR-BI.