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采用RT-PCR和RACE技术克隆了番茄LeWRKY1 cDNA(Genbank登录号为FJ654265)全长,用生物信息学的方法对其进行分析,预测其编码的蛋白的定位和功能。同时利用Northern杂交技术分析了JA(jasmonic acid)或SA(salicylic acid)刺激时LeWRKY1在野生型番茄和番茄JA不敏感突变体jail(JASMONATE-INSENSITIVE1)及番茄JA生物合成突变体spr2(prosystemin-mediated responses2)中的表达情况。LeWRKY1 cDNA序列全长1 740 bp,阅读框1 083 bp。生物信息学分析表明LeWRKY1含有1个WRKY结构域和1个C_2H_2型锌指结构。LeWRKY1可能是一种定位于细胞核的DNA结合蛋白,并可能具有转录调控、信号传导功能。表达分析推测LeWRKY1的表达是JA依赖而非SA依赖性的,且LeWRKY1的表达不依赖于生物体内JA的从头合成。
The full-length cDNA of LeWRKY1 (GenBank accession No. FJ654265) was cloned by RT-PCR and RACE techniques and analyzed by bioinformatics methods to predict the location and function of the encoded protein. Northern blotting was also used to analyze the effect of LeWRKY1 on JA (JASMONATE-INSENSITIVE1) and tomato JA biosynthesis mutant spr2 (prosystemin-mediated) in JA (jasmonic acid) or SA (salicylic acid) responses2). The full length of LeWRKY1 cDNA is 1 740 bp and the reading frame is 1 083 bp. Bioinformatics analysis showed that LeWRKY1 contains one WRKY domain and one C_2H_2-type zinc finger. LeWRKY1 may be a DNA-binding protein localized in the nucleus and may have transcriptional regulation and signaling functions. Expression analysis speculated that the expression of LeWRKY1 is JA-dependent rather than SA-dependent, and that the expression of LeWRKY1 does not depend on de novo synthesis of JA in vivo.