番茄DEAD-box RNA解旋酶基因SlDEAH1的克隆及表达分析  

Cloning and Expression Analysis of a DEAD-box RNA helicases Gene, SlDEAH1 in Solanum lycopersicum

作　　者：张建苓[1] 陈国平[1] 朱明库[1] 涂昀[1] 胡功铃[1] 胡宗利[1] 

机构地区：[1]重庆大学生物工程学院,重庆400044

出　　处：《植物研究》2014年第1期44-52,共9页Bulletin of Botanical Research

基　　金：重庆市自然科学基金(CSTC;2011BB1068);高等学校博士学科点专项科研基金(20100191110031);中央高校基本科研业务项目(CDJXS11230018)

摘　　要：DEAD-box RNA解旋酶是一种特殊的RNA分子伴侣,参与了RNA代谢,包括前体RNA剪接、核糖体合成、RNA降解以及基因表达,并对植物的发育和抗性等也具有重要作用。根据已报道的拟南芥DEAD-box蛋白,通过同源比对,在NCBI据库中筛选得到一个DEAD-box RNA解旋酶同源蛋白,命名为SlDEAH1,并根据其基因序列设计特异引物,应用RT-PCR方法从野生型番茄(Solanum lycopersicum)AC++中克隆得到了该基因的全长编码区序列。利用生物学网站、软件及实时荧光定量PCR方法,对其进行生物信息学、表达模式、胁迫及激素处理分析。结果表明:SlDEAH1包括2 073 bp的开放阅读框,编码690个氨基酸残基,其编码蛋白有9个保守结构基序,其所涉及到的ATP结合、ATP水解及RNA结合等功能对于解旋酶活性是至关重要的;表达模式分析表明SlDEAH1基因可能在野生型番茄萼片、叶片发育及果实成熟方面起到重要作用;高温、低温、脱水、伤害、盐胁迫不同程度的诱导了SlDEAH1的表达,但在根中该基因的表达受盐胁迫抑制;ABA、ACC、IAA、GA3、MeJA和ZT均不同程度诱导了SlDEAH1的表达,其中ABA诱导效应最为明显。这些结果为进一步研究SlDEAH1在番茄发育和胁迫响应中的功能奠定了基础。DEAD-box RNA helicases, which function as the RNA chaperones, are involved in RNA metabolism, including pre-mRNA splicing, ribosome biogenesis, RNA degradation and gene expression. In addition, they also play important roles in plant development and stress tolerance. A DEAD-box homologous protein was selected from the NCBI database by means of program of homologous alignment according to the published DEAD-box protein in Arabidopsis thaliana and maize and named S1DEAH1, then the specific primers, which being used to amplify the cDNA sequences of SIDEAH1 from wild type tomato AC^＋＋ , were designed according to the DNA corresponding sequences. Bioinformatics analysis, expression pattern, stress and hormone treatments were used to study the DEAD-box gene SIDEAH1. The results showed that the open reading frame of SIDEAH1 was 2 073 bp, encoding 690 amino acid residues. In the protein of SIDEAH1, 9 conserved motifs were found to be essential for the helicase activities such as ATP binding, ATP hydrolysis and RNA binding. RT-PCR expression pattern analysis showed that S1DEAH1 may play important roles in sepal and leaf development and fruit ripening. High temperature, low temperature, dehydration, wounding and salt stress induced the expression of SIDEAH1 in different degrees in leaves, but its expression in roots was obviously inhibited by salt stress. Furthermore, the expression of SIDEAH1 was also induced in different degrees by ABA, ACC, IAA, GA3, MeJA and ZT, and the inductive effect of ABA was the most obvious. These results lay a foundation for further study on the physiological function of SIDEAH1 in the development and stress response of Solanum lycopersicum.

关 键 词：番茄 DEAD-BOX RNA解旋酶 SlDEAH1 胁迫 激素 RT-PCR分析 

分 类 号：Q81[生物学—生物工程] S641.2[农业科学—蔬菜学]