关键词: pMD18-T-ambp, 克隆载体, 逆转录聚合酶链反应

Abstract: To obtain the cDNA sequence encoding human α₁-microglobulin/bikunin precursor (AMBP),and construct its recombinant cloning vector. Methods RNA was extracted from human embryo tissues of liver,kidney and pancreas with Trizol reagent.The sequence of human AMBP cDNA was cloned by RT-RCR from those RNAs,and inserted to pMD18-T vector.The recombinant vector pMD18-T-ambp was identified by restriction endonucleases SalⅠ/ EcoRⅠand DNA sequencing.Results The ambp gene fragment was obtained by RT-PCR only from liver RNA,but not from kidney and pancreas.A DNA fragment of 1 098 bp,consistent with the size of ambp gene fragment,was generated by digesting the recombinant plasmid with the restriction enzymes SalⅠ/EcoRⅠ.The results of sequence analysis showed that the constructed AMBP cDNA sequence was identical to the published one in GenBank. Conclusion The human AMBP cDNA is obtained successfully and the recombinant cloning vector pMD18-T-ambp is constructed correctly.

Key words: pMD18-T-ambp, cloning vectors, reverse transcriptase polymerase chain reaction