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丹参BTF3基因的克隆及生物信息学分析

Cloning and Bioinformatics Analysis of OsMsr3 Gene from Salvia miltiorrhiza Bunge

  • 摘要: 采用PCR及RT-PCR技术分别从丹参基因组DNA和cDNA水平克隆了通用转录因子3(Basic Transcription Factor3,BTF3)基因,运用生物信息学方法对该基因编码氨基酸序列的组成成分、疏水性/亲水性、翻译后修饰、蛋白质高级结构等进行预测,利用实时荧光定量PCR检测丹参不同器官以及脱落酸和干旱诱导下BTF3的表达情况。结果表明:丹参BTF3基因全长为1510 bp,包含5个外显子和4个内含子;cDNA全长794 bp,编码蛋白整体表现为亲水性,二级结构主要由α-螺旋、不规则卷曲组成,具有NAC结构域,与其他植物的BTF3在序列组成、结构及活性位点上有高度的相似性。丹参BTF3基因在根、茎、叶中均有不同程度的表达,表达丰度为:根>茎>叶;ABA诱导该基因表达量24 h内没有显著的变化;干旱胁迫下该基因的表达受到抑制。

     

    Abstract: A Basic transcription factor 3(OsMsr3) gene from Salvia miltiorrhiza Bunge(OsMsr3) was cloned by PCR and RT-PCR,named OsMsr3.The deduced amino acid sequence of SmOsMsr3 were analyzed by the tools of bioinformatics in the following aspects:the chemical composition,hydrophobicity or hydrophilicity,post-translational modification,structural domain and the advanced structure of deduced amino acid sequence.The expressions of SmOsMsr3 in different organs and under stress treatments were detected by real-time fluorescent quantitative PCR.The results showed that the genomic DNA of SmOsMsr3 was 1510 bp long,which contained 5 exons and 4 introns,while its cDNA was 794 bp.The deduced protein were hydrophilic.The second structure prediction indicated α-helix and random coil were the major motifs and NAC domain was included.The sequence composition,structure,and activity sites were highly similar with other BTF3 proteins in plants.The expression of OsMsr3 was the highest in root,followed by that in stem,and the least in leaf.Obvious changes of the expression of OsMsr3 were not Abserved during 24 h treated by ABA,while under the drought stress,the expression was inhibited at first and then increased to control level.

     

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