芜菁BrrLOX 7基因克隆、表达及生物信息学分析

Cloning,expression and bioinformatics analysis of BrrLOX7 gene in turnip

脂氧合酶(lipoxygenase,EC 1.13.11.12;简称LOX),是一类由非血红素铁组成的脂肪酸双加氧酶,在植物的生长发育和环境胁迫响应中发挥着重要作用,本研究以4片真叶期芜菁幼苗为材料,利用同源克隆技术从芜菁中克隆BrrLOX 7基因。利用生物信息学方法分析BrrLOX 7基因序列。使用荧光定量PCR技术分析非生物胁迫下BrrLOX 7基因表达模式。结果显示,克隆的BrrLOX 7基因CDS长度为2715 bp,编码904个氨基酸,编码的蛋白质分子量为103.10 ku,无跨膜结构和信号肽。亚细胞定位预测显示,该基因编码的蛋白位于叶绿体中,该蛋白与白菜和萝卜的序列同源性和遗传距离最近。不同胁迫下的表达模式显示,BrrLOX 7基因参与对4种胁迫(盐胁迫、冷胁迫、热胁迫、干旱胁迫)的响应。研究结果将为BrrLOX 7基因的功能研究奠定基础。

Lipoxygenase(EC 1.13.11.12;LOX)is a class of fatty acid dioxygenases composed of non-heme iron,which plays an important role in plant growth and development and environmental stress response.In this study,the BrrLOX 7 gene was cloned from turnip using homologous cloning techniques.The sequence of BrrLOX 7 gene was analyzed by bioinformatics.The expression pattern of BrrLOX 7 gene under abiotic stress was analyzed using fluorescence quantitative PCR.The results showed that the CDS length of cloned BrrLOX 7 gene was 2715 bp,it encoded 904 amino acids,protein encoded by BrrLOX 7 gene had a relative molecular weight of 103.10 ku and had no transmembrane structure or signal peptide.Subcellular localization prediction showed that the protein encoded by this gene was located in chloroplasts,and the protein had the closest sequence homology and genetic distance to Chinese cabbage and radish.Expression patterns under different stresses showed that the BrrLOX 7 gene was involved in response to four stresses(salt stress,cold stress,heat stress and drought stress).The results of this study will provide a basis for functional studies of the BrrLOX 7 gene.