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

作　　者：刘军梅[1] 李昊[1] 崔东清[1] 叶梅霞[1] 张志毅[1] 安新民[1] 

机构地区：[1]北京林业大学林木育种国家工程实验室、林木花卉遗传育种教育部重点实验室、国家林业局树木花卉育种与生物工程重点开放实验室,北京市100083

出　　处：《北京林业大学学报》2011年第4期55-62,共8页Journal of Beijing Forestry University

基　　金：林业公益性行业科研专项(201004009);国家自然科学基金项目(30571511);“863”国家高技术研究发展计划项目(2009AA10Z107)

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。To understand the genetic and molecular mechanisms during floral development in Populus tomentosa, we isolated a full-length cDNA sequence of PtlabA-likel 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-likel. 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-likel 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-likel. 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-likel was constructed. All of these would provide the groundwork for understanding the function of PtlabA-likel gene.

关 键 词：毛白杨 labA—like 克隆 表达 

分 类 号：S718.46[农业科学—林学]