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采用Affymetrix60K水稻基因表达芯片系统分析了水稻培矮64S(Oryza sativa ssp.indica)在低温、高温和干旱条件下不同生育时期的叶片与穗全基因组的转录丰度,筛选出一个类糖基转移酶基因OsCrGtl(Oryza sativ a cold responsive glycosyltransferase-like gene,GenBank登录号为FJ828672),该基因主要受低温诱导且在各生育时期的表达量均显著上调。用实时定量RT-PCR方法对该基因的在逆境条件下的转录丰度进行了验证分析,也得到与基因芯片分析基本相同的结果。以培矮64SmRNA为模板,通过RT-PCR方法扩增,得到包含其完整开放阅读框的cDNA克隆。根据其开放阅读框的核苷酸序列推测,OsCrGtl编码一个包含462个氨基酸残基的蛋白质,理论分子量为50.327kD,等电点为5.35。查询蛋白质数据库,得知该基因编码的蛋白质与籼稻(indica)、粳稻(japonica)、玉米(Zea mays)及拟南芥(Arabidopsis thaliana)编码的糖基转移酶家族蛋白相似,且包含糖基转移酶结构域。对其推测的启动子区域核苷酸序列进行分析,找到9个可能与逆境应答有关的顺式作用元件。
Affymetrix 60K rice gene expression microarray was used to analyze the transcriptional abundance of the whole genome of rice and leaves at different growth stages under low temperature, high temperature and drought conditions in Oryza sativa ssp. Indica. A glycosyltransferase The gene OsCrGtl (Oryza sativ a cold responsive glycosyltransferase-like gene, GenBank accession number FJ828672), the gene was mainly induced by low temperature and at each growth stage expression were significantly increased. The transcriptional abundance of this gene under adverse conditions was verified by real-time quantitative RT-PCR, and the same results as the gene chip analysis were also obtained. Using Pei-Pei 64S mRNA as a template, the cDNA clone containing its complete open reading frame was amplified by RT-PCR. According to the nucleotide sequence of its open reading frame, OsCrGtl encoded a 462 amino acid protein with a theoretical molecular weight of 50.327 kD and an isoelectric point of 5.35. The protein database was searched and the protein encoded by this gene was found to be similar to the glycosyltransferase family of proteins encoded by indica, japonica, maize (Zea mays) and Arabidopsis thaliana and contained glycosyltransferases Enzyme domain. The deduced nucleotide sequences of the promoter regions were analyzed to find nine cis-acting elements that might be related to adversity responses.