利用Gateway技术快速构建棉花GhANS基因植物表达载体

Construction of Plant Expression Vector for GhANS Gene of Cotton Using Gateway Technology

【目的】为了进一步探讨GhANS(花色素合成酶)基因的上调表达对彩色棉纤维色素合成的影响,构建了棉花GhANS基因的植物表达载体。【方法】利用Gateway技术的同源重组原理,将GhANS基因通过BP反应和LR反应分别重组到植物过表达载体pGWB17与干扰表达载体pANDA35HK中。【结果】经PCR和酶切鉴定,PCR扩增片段与酶切片段大小与预期结果相一致。【结论】基于Gateway技术的棉花GhANS基因的过表达载体和干扰表达载体已构建成功,为进一步研究该基因的功能及棉花的遗传转化奠定了基础。

[ Objective ] The purpose of this project is to construct cotton GhANS gene of plant expression vector in order to explore the effects of up - regulated expression of synthetase gene of flower pigments （GhANS） in cotton fiber pigment synthesis. [ Method] GhANS gene was restructured to the overexpression vec- tor pGWB17 and interference expression vector pANDA35HK via BP and LR reaction respectively based on the homologous recombination of Gateway technology. [ Result] The validation by PCR and restriction enzymes, segments of enzyme fragment and PCR products size were consistent with the expected results. [ Conclusion ] Based on the Gateway technology of cotton GhANS gene over - expression vector and interference expression vector has been successfully constructed, which has laid a foundation for further study of the function of genes and genetic transformation of cotton.