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    刘军梅, 李昊, 崔东清, 叶梅霞, 宋体, 张志毅, 安新民, . 毛白杨labA-like1基因的克隆与表达特性及其表达载体的构建[J]. 北京林业大学学报, 2011, 33(4): 55-62.
    引用本文: 刘军梅, 李昊, 崔东清, 叶梅霞, 宋体, 张志毅, 安新民, . 毛白杨labA-like1基因的克隆与表达特性及其表达载体的构建[J]. 北京林业大学学报, 2011, 33(4): 55-62.
    LIU Jun-mei, LI Hao, CUI Dong-qing, YE Mei-xia, SPAN lang, US style, pt solid, solid windowtext, ZHANG Zhi-yi, AN Xin-min. Isolation labA-like1 gene from Populus tomentosa: expression of PtlabA-like1 in P. tomentosa and construction of expression vector[J]. Journal of Beijing Forestry University, 2011, 33(4): 55-62.
    Citation: LIU Jun-mei, LI Hao, CUI Dong-qing, YE Mei-xia, SPAN lang, US style, pt solid, solid windowtext, ZHANG Zhi-yi, AN Xin-min. Isolation labA-like1 gene from Populus tomentosa: expression of PtlabA-like1 in P. tomentosa and construction of expression vector[J]. Journal of Beijing Forestry University, 2011, 33(4): 55-62.

    毛白杨labA-like1基因的克隆与表达特性及其表达载体的构建

    Isolation labA-like1 gene from Populus tomentosa: expression of PtlabA-like1 in P. tomentosa and construction of expression vector

    • 摘要: 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Conserved Domains Database (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-like superfamily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。

       

      Abstract: To understand the genetic and molecular mechanisms during floral development in Populus tomentosa, we isolated a full-length cDNA sequence of PtlabA-like1 gene from female floral bud cDNA library of P. tomentosa, with the method of bacteriophage absorption by filter paper-PCR. A conserved LabA-like superfamily domain was found via searching Conserved Domains Database. Therefore, it was given by the name PtlabA-like1. A phylogenetic tree was constructed using BioEdit and MEGA3.1 softwares to depict its evolutionary relationship with other 9 members comprising the same domain that retrieved via BLAST at JGI. The expression patterns of LabA-like superfamily members in tissues of P. tomentosa were estimated by RT-PCR. Results revealed that PtlabA-like1 and PtlabA-like6 had similar expression domains at stems, young leaves, mature leaves, male and female flower buds, but a higher abundance was especially shown in PtlabA-like1. PtlabA-like8 was detected with a low level at root only. Other members which had different abundances had a constitutive expression pattern at all of the six tissues. Furthermore, constitutive expression vector carrying 35S::PtlabA-like1 was constructed. All of these would provide the groundwork for understanding the function of PtlabA-like1 gene.

       

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