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利用RT-PCR技术从海岛棉品种新海21中克隆了1个WRKY基因,在GenBank数据库中比对发现其与陆地棉GhWRKY32的序列高度同源,因此命名为GbWRKY32。该基因ORF为1 077 bp,编码358氨基酸,预测分子量为39.288 k D,等电点为5.02。序列分析表明该基因含有一个WRKY保守结构域,锌指结构为:C-X5-C-X23-H-X1-H,属于WRKY转录因子家族Ⅱ类D组成员。GbWRKY32蛋白为疏水性蛋白,不具有跨膜区和信号肽结构。GbWRKY32转录因子不具有转录自激活活性。本研究成功构建GbWRKY32基因的植物表达载体并转入根癌农杆菌EHA105菌株中,这有助于该基因功能的后续研究。
A WRKY gene was cloned from the island cotton variety Xinhai21 by RT-PCR and was found to be highly homologous to GhWRKY32 in GenBank. Therefore, it was named GbWRKY32. The ORF of this gene was 1 077 bp, encoding 358 amino acids with a predicted molecular mass of 39.288 kD and an isoelectric point of 5.02. Sequence analysis showed that the gene contains a WRKY conserved domain. The zinc finger structure is C-X5-C-X23-H-X1-H, which belongs to class D group of WRKY transcription factor family. The GbWRKY32 protein is a hydrophobic protein that does not have a transmembrane domain and signal peptide structure. GbWRKY32 transcription factor does not have transactivation activity. In this study, the plant expression vector of GbWRKY32 gene was successfully constructed and transformed into Agrobacterium tumefaciens strain EHA105, which is helpful for the subsequent study of the function of this gene.