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为了探究非生物胁迫下甜瓜N–乙酰氨基葡萄糖基转移酶基因(CmGnT)的作用机制,根据甜瓜cDNA序列设计引物,利用RT-PCR技术克隆得到CmGnT。生物信息学分析表明,CmGnT的cDNA全长为2 193 bp,开放阅读框(ORF)为1 179 bp,编码392个氨基酸。CmGnT蛋白分子量约为46 kD,理论等电点(pI)为7.93。蛋白结构分析表明,CmGnT含有β–1,4–N–乙酰氨基葡萄糖转移酶结构域,属于糖基转移酶17家族。该基因编码的蛋白有1个强的跨膜螺旋结构。进化树分析表明,CmGnT氨基酸序列与黄瓜CsGnT(登录号:XP_004135275.1)亲缘关系最近,同源性为99.74%,与西瓜Cs Gn T的同源性为97.70%。实时荧光定量PCR分析结果表明,在NaCl、PEG、H_2O_2和ABA等非生物胁迫下的甜瓜叶片中CmGnT都显著上调表达,表明CmGnT受非生物胁迫诱导,推测其在甜瓜响09应非生物胁迫过程中起到重要作用。
In order to investigate the mechanism of CmGnT in melon under abiotic stress, primers were designed according to the cDNA sequence of melon, and CmGnT was cloned by RT-PCR. Bioinformatics analysis showed that the full-length cDNA of CmGnT was 2 193 bp and the open reading frame (ORF) was 1 179 bp, encoding 392 amino acids. The molecular weight of CmGnT protein is about 46 kD and its theoretical isoelectric point (pI) is 7.93. Protein structure analysis showed that CmGnT contains β-1,4-N-acetylglucosaminyltransferase domain, belonging to glycosyltransferase 17 family. The gene encodes a protein that has a strong transmembrane helix. Phylogenetic tree analysis showed that the amino acid sequence of CmGnT was the closest to cucumber CsGnT (accession number XP_004135275.1), with a homology of 99.74% and a homology of 97.70% with CsGnT of watermelon. The results of real-time PCR showed that CmGnT was up-regulated in the leaves of melon under abiotic stress, such as NaCl, PEG, H_2O_2 and ABA, which indicated that CmGnT was induced by abiotic stress. Play an important role.