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运用DNA重组技术,将用RT-PCR方法克隆的大鼠肾皮质受体相关蛋白319个氨基酸的核苷酸序列导入载体pBluescript和表达载体pGEX中,分别构建了重组质粒pBSK9RAP和pGEX-9RH7,转化入大肠杆菌XL1-Blue和DH5α中,经IPTG、x-gal和抗生素筛选阳性克隆,扩增后,重组质粒用限制性内切酶图谱分析及DNA测序分析,证明插入子插入方向正确,表达阅读框架正确。
The nucleotide sequence of 319 amino acids of rat renal cortex receptor-related protein cloned by RT-PCR was introduced into vector pBluescript and expression vector pGEX by using DNA recombination technology. Recombinant plasmids pBSK9RAP and pGEX-9RH7 were constructed and transformed respectively The recombinant plasmids were screened by IPTG, x-gal and antibiotics. After amplification, the recombinant plasmids were confirmed by restriction enzyme analysis and DNA sequencing. The framework is correct.