水稻花粉特异性启动子的克隆与表达

CLONING AND TRANSIENT EXPRESSION OF THE PROMOTER OF POLLEN-SPECIFIC GENE IN RICE (Oryza sativa L.)

利用PCR技术 ,从水稻幼苗总DNA中扩增并克隆了一段DNA片段 .将该片段与报告基因GUS基因融合 ,构成植物表达载体 .经农杆菌转化烟草萌发花粉 ,共培养约 2 4h后 ,作瞬时表达检测 .结果表明 ,该克隆片段具有启动子功能 .DNA序列分析表明 ,该克隆片段长度为 5 2 6bp ,含有TATA盒及CAAT盒 .与原基因(PSI)启动子序列比较 ,同源性为 5 2 % 。

The specific fragment was amplified and cloned from Oryza sativa L. cv Yizhao 204 by polymerase chain reaction (PCR) with the primers designed on the upsteam regulatory region of rice pollen specific gene (PS1). The fragment was orientedly fused to the reporter gene β glucuronidase (gus A) to form the expression vector. After co cultured with Agrobacterum tumefaciens carring the vector for about 24h, histochemical GUS assays indicated that the cloned fragment drived the GUS gene to express in mature and / or germinating pollen of tobacco (Nicotiana tobacum). Sequence analysis of the fragment showed that it has 526bp in length and contains TATA box and CAAT box. Comparison of this nucleotide sequence with that of published promoter region of PS1 revealed 52% of sequence homology and sharing some putative cis regulatory element motifs.