水稻胚乳特异性启动子Gt1的克隆及其功能验证

Cloning of a Rice Endosperm-specific Promoter Gt 1 and Its Functional Verification

以水稻品种“密阳 4 6”的 DNA为模板 ,用 PCR方法从水稻谷蛋白基因 Gt1上游序列扩增出特异性条带 ,并克隆出种子特异性启动子。经鉴定 ,它的长度为 92 9bp,与 Takaiwa(1987)报道的序列仅有 12个核苷酸的差异 ,已知的功能部位序列没有发生改变。用它构建了带动报告基因 Gus表达的双元载体 ,并用农杆菌介导法转化水稻得到了转基因植株。对 Gus基因表达检测结果表明 ,由该启动子序列引导 Gus基因能在胚乳中表达 ,而其它组织中都未表达 。

A pair of specific primer was designed according to the published sequence of the 5′ upstream sequence of rice glutelin gene Gt1 . A 929 bp sequence was amplified from genomic DNA template of an Indica rice variety 'Miyang 46' by PCR. Sequence analysis showed 12 bp difference between the cloned and the published sequences. No difference was found in known functional regions. The cloned promoter has been used in construction of Agrobacterium binary vectors with Gus reporter gene, and several transformed plants were obtained through Agrobacterium mediated transformation. Gus activity in various tissues of transformed plants was examined and the results showed that Gus gene directed by the promoter sequence of rice glutelin gene Gt 1 was tissue specifically expressed in endosperm, in contrast to the expression pattern of Gus gene directed by 35S promoter, which was expressed in all the tissues tested.