水稻Rubisco小亚基启动子的克隆及光诱导表达载体的构建

高效特异表达的启动子往往是转基因研究的关键因素。Rubisco小亚基启动子具有光诱导性、组织特异性和高表达的特性,可利用该启动子为转基因研究服务。本文以水稻（Oryza sativa）中花11叶片为材料,根据GenBank所报道的水稻Rubisco小亚基启动子序列,设计特异引物从水稻基因组DNA中扩增得到长度约1600bp的DNA片段,将该片段连接至T载体pMD18-TSimple,测序结果表明该片段序列与GenBank报道序列一致为100%。Plant CARE序列分析表明,该启动子具有12个与光诱导表达相关的元件。为构建光诱导表达载体,将该启动子和植物表达载体pCactF分别以KpnⅠ和XbaⅠ酶切后连接。光诱导表达载体的构建为进一步研究基因功能及利用光诱导表达载体改良作物品质奠定基础。

Cloning of the Promoter of the Rubisco Small Subunit in Rice and Construction of Light-inducible Expression Vector

A high efficiency and specific expression promoter ususlly is one of the key factors in transgenic plant research. The promoter of Rubisco small subunit （rbcS） has light-inducible, tissue-specific and high expression characteristics, which could be used in transgenic plant research. A pair of specific primers was designed according to the DNA sequence of the promoter of rbcS of rice, and a DNA fragment （about 1 600 bp） was amplified from rice zhonghua 11 genome DNA by PCR using the specific primers. The DNA fragment was then cloned into pMD18-T Simple, the sequencing result showed that the sequence was identical to the sequence in GenBank with 100% identity. In order to construct light-inducible expression vector, the rbcS promoter was cloned into plant expression vector pCactF after being digested with KpnⅠ and XbaⅠ, which provided scientific reference for further study gene function and use of the vector to improve crop quality.