毛白杨labA-1ikel基因的克隆与表达特性及其表达载体的构建

利用滤纸吸附噬菌体一PCR法，从毛白杨花芽cDNA文库中分离克隆了PtlabA-1ikel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—likel。经BL...

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

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.