ZAK-β蛋白激酶基因真核表达载体的构建

Construction of ZAK-β Eukaryotic Expression Vector

目的将人源ZAK-β基因定向克隆入MSCV-IRES-GFP质粒,构建真核表达载体p MSCV-IRES-GFP-ZAK-β.方法以肺癌A549细胞的c DNA为模板,通过PCR得到ZAK-β特异性基因产物,克隆入真核表达载体p MSCV-IRES-GFP中,获得重组真核表达载体p MSCV-IRES-GFP-ZAK-β,测序验证.结果扩增了ZAK-β基因,经双酶切、测序鉴定证实目的基因克隆到真核表达载体p MSCV-IRES-GFP中,测序结果与预测完全一致.结论成功构建了ZAK-β逆转录病毒表达载体,为进一步研究ZAK蛋白激酶的功能奠定了基础.

Objective To construct a recombinant retroviral vector containing human ZAK-βcDNA. Methods ZAK-β cDNA was amplified from lung cancer A459 cells by RT-PCR with ZAK-β-specific primers. PCR product was cloned into eukaryotic expression vector pMSCV-IRES-GFP, and then sequenced. Results ZAK- β cDNA was successfully amplified by RT-PCR. Expression vector pMSCV-IRES-GFP-ZAK- was also successfully obtained and confirmed. Conclusion The recombinant retroviral vector bearing human ZAK-β cDNA was successfully constructed, which forms the basis for the further studies on ZAK-β function.