人黑素瘤细胞A375 MIA/CD-RAP基因的克隆及其鉴定

Clone of Human MIA/CD-RAP cDNA and Its Authentication

目的为了研究人黑素瘤M IA/CD-RAP基因的生物学功能,克隆M IA/CD-RAP的cDNA,构建表达载体后,对其进行酶切和DNA测序鉴定。方法用RT-PCR从培养的人黑素瘤细胞A375中扩增M IA/CD-RAP cDNA,将其克隆到载体pcDNA3.1-MycH is上,经酶切及PCR扩增鉴定和测序。结果酶切电泳和DNA测序结果表明,成功地克隆了人黑素瘤M IA/CD-RAP cDNA。结论人黑素瘤M IA/CD-RAP cDNA的克隆,可为进一步研究其生物学功能奠定基础。

Objective To study the functional role of MIA/CD-RAP in melanoma, MIA/CD-RAP cDNA was cloned and authenticated. Methods MIA/CD-RAP cDNA of human melanoma was amplified by RT-PCR,cloned into vector pcDNA3.1-MycHis,then amplified by PCR,authenticated by digestive enzymes and sequenced. Results Authentication,PCR and DNA sequencing showed the recombinant plasmid of human MIA/CD-RAP was successfully constructed. Conclusion Cloning of human MIA/ CD-RAP cDNA facilitated further investigation of its biological function in melanoma.