姚彩萍, 王宗阳, 蔡秀玲, 张栋, 张景六, 洪孟民
将水稻翻译起始点(ATG)5^上游3.4 kb(-2118-＋1291 bp)片段经外切核酸酶Exo III部分酶解,得到一系列5^端缺失的片段。将这些缺失片段分别与gus基因编码区连接,构建成融合质粒,经PEG介导引入水稻原生质体,26℃培养48h后,定量测定GUS酶活力,并以同时导入的由35S启动子指导的荧光素酶(LUC)基因表达的酶活力作为内对照。结果表明,GUS酶活性随5^上游调控区长度的减少而逐渐减弱。由-861 bp缺失至-640 bp时,gus基因表达水平有较明显的降低,推测在该区域中可能存在一个顺式作用元件区。
Effects of 5^ upstream region deletion on rice waxy gene expression
Yao Caiping, Wang Zongyang, Cai Xiuling, Zhang Dong, Zhang Jingliu, Hong Mengmin
Shanghai Institute of Plant Physiology, The Chinese Academy of Sciences. Shanghai 200032
To identify the cis-acting element (s) within the 5^ upstream region of the rice waxy gene, a series of deletion mutants was constructed by using Exo III to digest the waxy promoter from 5^ end. Each of the chimeric genes consisting of these deleted sequences and coding sequence of beta-glucuronidase (GUS) gene was respectively introduced into rice protoplasts mediated by PEG and the expression of the chimeric gene was examined after incubation of these protoplasts for 48 h at 26 ℃. The luciferase (LUC) gene was co-introduced in each experiment, and its expression evaluated as an internal control. The data showed that the GUS activity decreased as the 5^ upstream region was shortened. When the waxy promoter was deleted from -861 bp to -640 bp, the GUS relative activity dramatically decreased. this result implicates that a cis-acting element located between -861 bp and -640 bp may play a regulatory role in the expression of the rice waxy gene.
姚彩萍, 王宗阳, 蔡秀玲, 张栋, 张景六, 洪孟民. 水稻蜡质基因5^上游区缺失对基因表达的影响[J]. 生理学报 1996; 48 (4): .
Yao Caiping, Wang Zongyang, Cai Xiuling, Zhang Dong, Zhang Jingliu, Hong Mengmin. Effects of 5^ upstream region deletion on rice waxy gene expression. Acta Physiol Sin 1996; 48 (4): (in Chinese with English abstract).