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利用滤纸吸附噬菌体--PCR法,从毛白杨花芽cDNA文库中分离克隆了PtlabA-like1基因cDNA全长序列。在ConservedDomainsDatabase(CDD)中对该基因氨基酸保守结构域进行分析,发现其包含一个LabA-likesuperfamily结构域,故将其命名为PtlabA-like1。经BLAST分析,在毛果杨基因组数据库中检索到含有该保守结构域的其他9个家族成员。运用BioEdit和MEGA3.1软件,绘制了该基因家族表达蛋白的系统进化树。通过RT-PCR分析,揭示了该基因及其他家族成员在毛白杨中的组织差异表达模式:PtlabA-like1、PtlabA-like6呈现相似的组织表达模式,在茎、幼叶、成熟叶和雌雄花芽组织中均表达,但PtlabA-like1表达丰度较高;PtlabA-like8只在根部表达,且表达丰度较低;其他成员在根、茎、成熟叶、幼叶和雌雄花芽组织中均有表达,但是不同成员的表达丰度存在一定的差异。为进一步研究该基因的功能,构建了该基因的正义表达载体。
The full-length sequence of PtlabA-like1 cDNA was isolated and cloned from the cDNA library of Populus tomentosa by filter paper adsorption phage-PCR. The amino acid conserved domain of this gene was analyzed in ConservedDomainsDatabase (CDD) and found to contain a LabA-likesuperfamily domain, hence its name as PtlabA-like1. After BLAST analysis, nine other family members containing this conserved domain were found in P. sinicus genome database. Using BioEdit and MEGA3.1 software, the phylogenetic tree of protein expressed by this gene family was drawn. RT-PCR analysis revealed that the gene and other family members in the Populus tomentosa differential expression patterns: PtlabA-like1, PtlabA-like6 showed a similar pattern of tissue expression in stems, young leaves, mature leaves and male and female flower bud tissue While PtlabA-like8 was only expressed in the roots and had lower abundance. The other members were expressed in roots, stems, mature leaves, young leaves and male and female flower buds, However, there are some differences in the abundance of different members. To further study the function of this gene, we constructed a sense expression vector of this gene.