人再生肝脏蛋白磷酸酶-3基因的克隆与测序

Gene of Human Phosphatase of Regenerating Liver-3: Cloning and Sequencing

目的:从人结肠癌肝转移病灶标本中提取染色体RNA,PCR扩增及克隆人PRL-3基因。方法:收集人结肠癌肝转移标本,提取总RNA,逆转录为cDNA,PCR扩增PRL-3序列,A-T克隆于pMD-18T载体中,转化大肠杆菌JM109株,提取质粒,对重组质粒进行酶切与测序鉴定。结果:通过酶切、PCR与测序三种方法证实所获得的基因片断为PRL-3基因。结论:克隆到序列正确的人源性PRL-3基因,为实现PRL-3基因的高效表达及制备相应抗体奠定了基础。

Objective：To amplify and clone the coding sequence of phosphatasc of regenerating liver-3 （ PRL-3） from the human liver metastasis sample of colorcctal cancer. Methods：Total gcnomic RNA was extracted from human liver metastasis sample and reversely transcriptcd to eDNA. The coding sequence of PRL-3 was amplified by using PCR technique. Then the gcnc was linked to pMD18-T vector and transformed to E. coli JM109. The recombinant plasmid was identified by Barn H Ⅰ , Hind Ⅲ digestion and sequenced. Results： The genetic frcgmcnt was identified to be the gcnc of PRL-3 with enzyme digestion, PCR and sequencing techniques. Conclusion：The correct human PRL-3 gcne was successfully amplified, thus laying a foundation for highly effective expression and development of the coded protein and preparation of its antibody.