论文部分内容阅读
蔗糖磷酸合酶是植物中控制蔗糖合成过程中催化6-磷酸果糖转化为蔗糖的关键酶。为了研究该基因的表达特性,本实验采用RT-PCR方法从甜菜中克隆甜菜蔗糖磷酸合酶基因(BvSPS1)。BvSPS1开放阅读框为3138bp,编码1045个氨基酸。推测BvSPS1氨基酸序列跨膜区域位于第561～593位,与酿酒葡萄(Vitis vinifera)和烟草(Nicotiana tabacum)的序列同源性分别为75.45%和74.59%。利用半定量RT-PCR对BvSPS1进行组织特异性表达检测,结果表明该基因主要在主根及侧根表达,在叶和叶柄中表达较弱,酶活性分析结果与之相同。离体叶片在10%葡萄糖溶液中培养6～12h后,BvSPS1基因表达水平提高,在10%蔗糖中培养相同时间则无变化。
Sucrose phosphate synthase is a key enzyme in plants that controls the conversion of fructose-6-phosphate to sucrose during sucrose synthesis. In order to study the expression characteristics of this gene, we isolated the beet sucrose phosphate synthase gene (BvSPS1) from beet by RT-PCR. BvSPS1 open reading frame is 3138bp, encoding 1045 amino acids. The deduced amino acid sequence of BvSPS1 was located between the 561th and the 593th positions in the transmembrane region, and the homologies with the sequences of Vitis vinifera and Nicotiana tabacum were 75.45% and 74.59%, respectively. Tissue-specific expression of BvSPS1 was detected by semi-quantitative RT-PCR. The results showed that the gene was mainly expressed in main and lateral roots, but weakly expressed in leaves and petioles. The results of enzyme activity analysis were the same. After cultured in 10% glucose solution for 6 ~ 12 h, the expression level of BvSPS1 gene increased in vitro, and no change was observed in 10% sucrose at the same time.