大麦幼穗MLOC-14401基因编码区的同源克隆和序列分析

Homology Cloning and Analysis on the Coding Domain Sequence of MLOC-14401 Gene of Barley Immaturity Spike

【目的】编码区序列是基因作用机制、遗传多样性和进化关系等研究的重要资源。【方法】以盐城红茎大麦、青田红大麦、淳安六棱胭脂大麦和北青7号为材料,采用同源克隆技术分离、克隆MLOC-14401基因CDS序列,利用ORF Finder、BLAST和Clustal X等软件分析所克隆的CDS序列。【结果】MLOC-14401基因CDS序列全长1 248 bp、包含3个外显子、GC含量为62.82%,位于大麦3H染色体的609 892 778~609895 303 bp区间,起始密码(ATG)和终止密码序列(TAA)分别位于119 bp和1 366 bp位点。所编码多肽链包含415个氨基酸、2个MYB结构域,与普通小麦等9个物种的MYB基因编码多肽链具有不同程度的氨基酸序列相似性,保守序列位于105氨基酸与237氨基酸之间。【结论】MLOC-14401基因属大麦转录调控基因R2R3-MYB,所克隆的CDS序列对于大麦MYB基因作用机制等研究具有一定指导作用。

[Purpose]The DNA sequence of coding domain is an important resource for the studies including action mechanism, genetic diversity and evolutionary relationships of gene.[Method]In the present paper, 4 barley varieties, Yanchenghongjingdamai, Qingtianhongdamai, Chunanliulingyanzhidamai and Beiqing7 were used as trail materials, the homology cloning technology were used to clone the coding domain sequence（CDS） of gene MLOC-14401, and softs including ORF Finder,BLAST and Clustal X were used to analysis sequences in the coding domain of gene MLOC-14401.[ Result] The CDS of gene MLOC-14401 contains 1 248 bases, 3 exions and 62.8% GC bases, was located on the region between 609 892 778–609 895 303 bp of 3 H chromosome. In the CDS, start codon（ATG） and stop codon（TAA） were located at 119 bp locus and 1 366 bp loci, respectively. The polypeptide coded by the CDS contains 425 amino acids, 2 MYB domains, and shows the some similarity between the amino acid of polypeptide chain in 9 species including wheat, the conserved sequence of these polypeptide chains were located in region between 105 amino acid locus and 237 amino acid locus.[Conclusion]The gene, MLOC-14401, belongs to barley transcription regulatory gene R2 R3-MYB, the cloned CDS is useful for the studies including action mechanism of barley MYB gene.