桑树Gtpase Era基因的克隆及非生物胁迫表达分析

Zhao Weiguo
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摘要

Era(大肠杆菌ras样蛋白)形成了一个独特的GTPase家族,其功能是通过将细胞分裂与生长速率偶联来控制细胞周期。本文从桑树中克隆了GTPase Era mRNA的cDNA序列。序列分析显示其开放阅读框(ORF)长度为1275 bp,编码一个424个氨基酸的蛋白。GTPase Era基因具有Era结构域,属于FeoB_N超家族。根据不同种GTPase基因编码的氨基酸序列进行系统发育分析,桑与桑(Morus notabilis)、山核桃(Ipomoea nil)、辣椒(Capsicum annuum)、秋葵(Solanum pennellii)、烟叶(Nicotiana sylvestris)和烟草(Nicotiana tubacum)亲缘关系较近。采用实时荧光定量PCR (RT-qPCR)检测干旱、低温和盐胁迫下GTPase Era的表达谱。与正常生长环境相比,干旱、寒冷和盐胁迫处理下mRNA的表达量发生了显著变化,干旱胁迫第3天、低温胁迫第6天、盐胁迫第7天mRNA表达量达到最大值。这些数据提供了更好地了解桑树胁迫反应过程中信号转导的分子基础
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Molecular Cloning and Abiotic Stress Expression Analysis of Gtpase Era Gene in Mulberry (Morus Alba L.)
Era (E. coli RAS-like protein) forms a unique family of GTPase, its functions in cell cycle control by coupling cell division with growth rate. In the present paper, a cDNA sequence coding GTPase Era mRNA was cloned from mulberry. Sequence analysis showed that its open reading frame (ORF) is 1275 bp in length encoding a protein of 424 amino acids. The GTPase Era gene had era domain and belonged to the FeoB_N superfamily. Phylogenetic analysis based on the amino acid sequences encoded by the GTPase gene from various species showed that the mulberry was closely related to Morus notabilis, Ipomoea nil, Capsicum annuum, Solanum pennellii, Nicotiana sylvestris and Nicotiana tubacum. The expression patterns of GTPase Era treated under drought, low temperature and salt stresses were examined using real-time quantitative PCR (RT-qPCR). The expression level of the mRNA had a significant change under drought, cold and salt stress treatments compared to the normal growth environment with the maximum expression on third day for drought stress, the sixth day for the low temperature stress and the seventh day for the salt stress. These data provide a better understanding of the molecular basis of signal transduction during stress responses in mulberry trees
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