共表达红光荧光蛋白及发夹RNA载体的构建及鉴定

Construction and identification of vectors encoding both red fluorescent protein and shRNA

目的 :构建共表达红光荧光蛋白及短发夹状 RNA的重组真核载体。方法 :利用亚克隆、T- A克隆和PCR技术构建 pc DNA3.0 / Ds Red- U6 - sh GFR及 pc DNA3.0 / Ds Red- U6重组载体。结果 :成功构建上述两种真核重组表达载体。结论 :这种共表达载体的构建为进一步利用 RNAi技术研究真核细胞基因功能奠定了基础。

Objective To construct the recombinant vectors that coexpress red fluorescent protein and small hairpin RNA (shRNA). Methods The subcloning, T-A cloning, and PCR technique were used to construct the recombinant vectors,named pcDNA3.0/DsRed-U6-shGFP and pcDNA3.0/DsRed-U6. Results The recombinant expression vectors mentioned above were successfully constructed.Conclusion The successful construction of recombinant vectors can establish a feasibility to further explore the functions of specific genes in eukaryotic cells with RNAi technique