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烟草ABF转录因子基因的克隆与生物信息学分析

Clone and Bioinformatics Analysis of ABF Transcription Factor Gene from Nicotiana tabacum

  • 摘要: 为获得烟草AREB/ABF(ABA响应元件结合蛋白/ABRE结合因子)家族转录因子基因序列,进一步提高烟草的抗渗透胁迫能力,通过同源比对和RT-PCR技术,从烟草品种红花大金元中克隆到烟草AREB/ABF家族转录因子NtABF1(GenBank登录号为KF736849),NtABF2(GenBank登录号为KF736850)基因编码序列,并进一步利用生物信息学分析软件对这两个转录因子的蛋白理化性质、高级结构和生物学功能进行了预测。结果表明:NtABF1基因开放阅读框(Open Reading Frame,ORF)长999 bp,编码332个氨基酸,蛋白分子量36.45 kDa,等电点9.04;NtABF2基因ORF长1305 bp,编码433个氨基酸,蛋白分子量46.92kDa,等电点9.35。NtABF1和NtABF2均为含有BRLZ(Basic Region and Leucine Zipper)保守结构域的亲水蛋白,不含信号肽与跨膜结构。其高级结构均以α螺旋和无规则卷曲为主,β转角和延伸链含量较少。在亚细胞水平上,NtABF1和NtABF2均定位于细胞核。磷酸化位点分析表明,两蛋白均含有丝氨酸,苏氨酸和酪氨酸等激酶识别位点。进化分析和多序列比对结果表明,NtABF1与AtAREB3的进化关系最近,序列相似性达54.49%;NtABF2与SlAREB1,StABF1和StABF的亲缘关系较近,序列相似性分别为83.63%,83.52%和83%。功能预测结果显示,NtABF1和NtABF2均为转录调控因子。

     

    Abstract: To obtain the sequences of tobacco AREB/ABF(ABA responsive element binding proteins/ABRE binding factor) family transcription factor gene and improve tobacco's resistance to osmotic stress,the gene coding sequences of two tobacco AREB/ABF family transcription factors,NtABF1(GenBank accession number KF736849) and NtABF2(GenBank accession number KF736850),were cloned from tobacco cv.Honghuadajinyuan via sequence homology alignment and RT-PCR technique.The protein physicochemical properties,advanced structures and biological functions of the two transcription factors were predicted by bioinformatics analysis software.The results indicated that the Open Reading Frame(ORF) of NtABF1 gene was 999 bp in length,which encoded 332 amino acids with the protein molecular mass of 36.45 kDa and the isoelectric point of 9.04;the ORF of NtABF2 gene was 1305 bp in length,which encoded 433 amino acids with the protein molecular mass of 46.92 kDa and the isoelectric point of 9.35.Both NtABF1 and NtABF2 were hydrophilic proteins containing conservative structure domain Basic Region and Leucine Zipper(BRLZ),neither of them had signal peptide or transmembrane structure domain.Their advanced structure was mainly Alpha helix and random coil,the contents of Beta turn and extended strand were less.At subcellular level,both NtABF1 and NtABF2 localized in nucleus.Phosphorylation locus analysis indicated that the two proteins both contained recognition loci of Ser,Thr,Tyr kinases.The results of phylogenetic analysis and multiple sequence alignment showed that NtABF1 had closer genetic distance with AtAREB3 and their sequence similarity was 54.49%,NtABF2 had closer genetic distance with SlAREB1,StABF1 and StABF,and their sequence similarities were 83.63%,83.52% and 83%,respectively.Functional prediction results indicated that both NtABF1 and NtABF2 functioned as transcriptional regulators.

     

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