Hsa-miR-200a真核表达载体的构建及鉴定

Construction and identification of eukaryotie expression vector of hsa-miR-200

目的:通过构建及鉴定miR-200a的真核表达载体pcDNA3.1(+)-miR-200a,为进一步研究miR-200a的功能奠定实验基础。方法:RT-PCR扩增pre-miR-200a基因序列,连接于表达载体pcDNA3.1(+)中,对重组质粒双酶切鉴定并测序验证。结果:pcDNA3.1(+)-miR-200a真核表达载体经酶切及测序鉴定正确。结论:pcDNA3.1(+)-miR-200a真核表达载体构建成功,为下一步深入研究miR-200a的生物学功能和验证miR-200a的靶基因提供有效工具。

Objective： To construct and identify the eukaryotic expressiou vector pcDNA3.1 （ ＋ ） - miR - 200a of hsa - miR - 200a. It may lay a foundation for further studying the function of hsa - miR - 200a. Methods： Amplified by reverse transcription - polymerase chain reaction（ RT - PCR）, The pre - miR - 200a sequence was inserted into pcDNA3.1 （ ＋ ） vector. The recombinant plasmid of pcDNA3.1 （ ＋ ） - miR - 200a was confirmed by restriction endnuclease analysis as well as DNA sequencing. Results： The result of DNA sequencing demonstrated that eukaryotic vector pcDNA3.1 （ ＋ ） - miR- 200a was completely correct. Conclusion： The successful construction of eukaryotic expression vector of hsa - miR - 200a provided a useful tool for the study of biological function of hsa - miR - 200a and its potential target genes.