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本研究从已构建的苦瓜果实均一化文库筛选得到一个EIN3-like同源EST序列,结合RACE技术,克隆得到EIN3基因c DNA全长序列,命名为Mc EIL2(Gen Bank登录号:KF595122)。该基因全长2 122 bp,开放阅读框1 890 bp,编码629个氨基酸,预测该蛋白的分子量为71.58 k D,与黄瓜Cus EIN3、甜瓜Cm EIL1、苜蓿Mt EIL1、绿豆Vr EIL1和番茄Le EIL1的蛋白同源性分别为89.61%、78.37%、71.23%、69.09%和65.66%。Mc EIL2基因的编码蛋白亚细胞定位于细胞核,荧光定量分析表明Mc EIL2基因表达量在幼果期先强后弱,绿熟期最低,破色期表达量大幅增加至最高随后下降,推测该基因参与苦瓜果实成熟软化调控过程。本研究初步明确了Mc EIL2基因在苦瓜成熟过程表达模式,可为今后进一步揭示苦瓜果实成熟软化的分子机制奠定基础。
In this study, an EIN3-like homologous EST sequence was screened from the constructed homozygous library of bitter melon fruit, and the full-length cDNA of EIN3 gene was cloned by RACE technology and named as Mc EIL2 (Gen Bank accession number: KF595122). The gene was 2 122 bp in length and 1 890 bp in open reading frame (ORF) encoding 629 amino acids. The predicted molecular weight was 71.58 kD, which was close to those of cucumber Cus EIN3, melon Cm EIL1, alfalfa Mt EIL1, mung bean Vr EIL1 and tomato Le EIL1 The protein homologies were 89.61%, 78.37%, 71.23%, 69.09% and 65.66%, respectively. The expression of Mc EIL2 gene was localized in the nucleus. Fluorescence quantitative analysis showed that the expression level of Mc EIL2 gene was weaker than that in the early stage, the lowest in the green ripe stage, the highest in the breakage stage, and then decreased. Participate in bitter melon fruit ripening and softening process. This study initially identified the expression pattern of McEIL2 gene in the process of bitter melon maturation, which may lay the foundation for further revealing the molecular mechanism of bitter melon ripening and softening.