植物HSP70反义基因工程雄性不育表达载体的构建及鉴定

Construction and Identification of HSP70 Antisense RNA Expression Vector for Genetic Engineering Male Sterility in Plant

[目的]为更好地研究HSP70热激蛋白基因与植物雄性不育的关系。用PCR方法克隆了水稻花药特异表达启动子Osg6B,将其与设计合成的HSP70反义片段连接,构建了由水稻花药特异表达启动子Osg6B驱动的HSP70反义表达载体,并进行了PCR和酶切鉴定。[结果]克隆的Osg6B启动子序列与所发表的序列同源性高达97%,启动子区域的顺式调控元件完整;构建的由水稻花药特异表达启动子Osg6B驱动的HPS70反义表达载体通过了菌落PCR验证和表达载体值粒的酶切鉴定,载体构建成功。[结论]该表达载体的构建将为植物基因工程雄性不育的利用奠定基础。

[ Objective]In order to study the relation between the HSPTO gene and male sterility of plant further. [ Methods] Anther specific expression promoter Osg6B of rice was coloned by PCR then connected with HSP70 antisense fragment to construct HSP70 antisense expression vector. The expression vector was identified by PCR experiment and enzyme digestion. [ Rresult ] The sequence of coloned Osg6B promoter had 97 % homology to the published sequence, and the cis-regulatory element in pronloter area was integrated. HSP70 antisense expression vector driven by the promoter Osg6B was confired by colony PCR and enzyme digestion. [Conclusion]the construction of expression vector would lay solid foundation for utilization of genetic engineering male sterility of plant.