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基于我们最近获得的小桐子低温驯化转录组和数字基因表达谱数据,本工作研究了低温驯化条件下差异表达变化较大的非特异性脂质转移蛋白A基因Jcns LTPA。克隆到该基因的c DNA序列全长833 bp,开放阅读框长度513 bp,编码170个氨基酸,存在ATT_LTSS典型保守功能基序。其启动子区域中鉴定到了TATA框、CAAT框、CATA框、W框等顺式作用元件以及CRT/DRE低温响应元件。半定量RT-PCR分析表明,该基因在茎、根、叶中都有表达,以茎中表达量最高、且受低温诱导最显著。同时,酵母表达Jcns LTPA也提高了重组酵母菌的抗低温能力。这些结果充分说明了小桐子Jcns LTPA是与抗冷性密切相关的基因,可以用于小桐子的抗冷性遗传改良。
Based on our recently obtained cryogenic acclimation transcriptome and digital gene expression profiling data, we studied Jcns LTPA, a nonspecific lipid transfer protein A gene, with a large variation in expression under low temperature acclimation conditions. The cloned cDNA sequence of the gene was 833 bp in length and 513 bp in open reading frame, encoding a protein of 170 amino acids. A typical conserved motif of ATT_LTSS was found. Its promoter region identified TATA box, CAAT box, CATA box, W box and other cis-acting elements and CRT / DRE low temperature response element. Semi-quantitative RT-PCR analysis showed that the gene was expressed in stem, root and leaf, with the highest expression in stem and the most significant induction by low temperature. At the same time, yeast expression of Jcns LTPA also increased the anti-cryogenic ability of the recombinant yeast. These results fully demonstrate that Jatropha Jcns LTPA is a gene closely related to cold resistance and can be used for genetic improvement of chilling resistance in Jatropha curcas.