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为了解叶用莴苣热激蛋白LsHsp70-1707基因在高温下的作用机制,通过同源克隆及RACE技术克隆并获得叶用莴苣热激蛋白LsHsp70-1707基因全长cDNA序列,实时荧光定量PCR(qRT-PCR)分析基因在不同叶用莴苣品种中的表达量差异。该基因全长2 400bp(KP258179),开放阅读框为2 106bp,编码701个氨基酸,与西瓜(AAC03416.1)、牵牛花(ABZ04081.1)、黄瓜(CAA52149.1)、菠菜(AAB91471.1)等HSP70蛋白高度同源。实时荧光定量PCR结果表明:高温处理时,该基因在热敏品种中表达没有明显增加,随着处理时间的增加甚至表现为下调,而在耐热品种中,其表达上调。成功克隆得到叶用莴苣热激蛋白LsHsp70-1707基因,热胁迫下该基因在不同品种中的表达有一定的差异性,推测该基因可能与叶用莴苣耐热性相关。
In order to understand the mechanism of leaf heat-induced heat shock protein LsHsp70-1707 at high temperature, full-length cDNA sequence of lettuce heat shock protein LsHsp70-1707 was cloned by homologous cloning and RACE techniques. Real-time quantitative PCR (qRT -PCR) analysis of gene expression in different leaf lettuce varieties of expression differences. The full length of this gene was 2 400 bp (KP258179) with an open reading frame of 2 106 bp encoding a polypeptide of 701 amino acids. The full length of this gene was associated with AAC03416.1, ABZ04081.1, CAA52149.1 and AAB91471. 1) and other HSP70 protein highly homologous. The results of real-time PCR showed that the gene expression did not increase significantly in heat-sensitive varieties at high temperature, even down-regulated with the increase of treatment time, but up-regulated in heat-tolerant varieties. The gene of LsHsp70-1707 was successfully cloned from leaf lettuce heat-stressed protein, and the expression of LsHsp70-1707 was different under different heat stress. It is speculated that this gene may be related to the heat tolerance of leaf lettuce.