巨桉EgrWOX5基因的克隆、表达分析与超表达载体构建

Expression Analysis and Molecular Cloning of WOX5 in Eucalyptus grandis and Over-expression Vector Constructed

以巨桉(Eucalyptus grandis)EG5无性系为试验材料,通过RT-PCR技术从中克隆得到了1个巨桉WOX家族基因成员的CDS序列,命名为Egr WOX5(Gen Bank登录号为KF964019)。该基因全长为543 bp,编码181个氨基酸,具有WOX基因家族最保守的特征结构域-Homeobox domain(HD)结构域;其氨基酸序列与草本植物的拟南芥At WOX5基因和木本植物杨树Ptr WOX5基因的氨基酸序列有较高的相似性。组织表达分析结果显示,Egr WOX5基因特异性地在巨桉EG5的根组织中表达。这些结果间接或直接地说明Egr WOX5在巨桉根组织的发育过程中发挥重要作用。本研究还利用Gateway技术成功构建了Egr WOX5基因的超表达载体,为后续基因功能的进一步鉴定奠定基础。

A member of WOX gene family,Egr WOX5（Genbank Accession Number is KF964019） was cloned from Eucalyptus grandis using RT-PCR,which had the most conserved domain-Homeobox domain.The gene is 543 bp and encodes 181 amino acids.The deduced amino acid sequence of Egr WOX5 shares high similarity with At WOX5 from Arabidopsis thaliana and Ptr WOX5 from Populus trichocarpa.Expression analysis revealed its tissue-specific pattern in root.In conclusion,all the results could directly or indirectly prove Egr WOX5 play important roles in the development of root in E.grandis.The study also took advantage of Gateway technology and successfully constructed the over-expression vector of Egr WOX5,which would lay a foundation for further functional study of the gene.