Cloning of CaWRKY39 Gene in Capsicum annuum L.and Its Yeast-One-Hybrid pGADT7 Vector Construction
Pepper(Capsicum annuum L.)is one of the most important economic crops in China,but To-mato spotted wilt virus(TSWV)has caused huge losses in its production,so effective control strategies are ur-gently needed.Previous research found that a WRKY transcription factor CaWRKY39 was specifically highly expressed after TSWV infection,suggesting that it may play a critical role in pepper resistant to TSWV.In this study,the CaWRKY39 gene was cloned from leaves of Xiaoxin 19 pepper.The bioinformatics analysis results showed that the full length of CaWRKY39 CDS was 1 035 bp,which encoded a stable hydrophobic protein composed of 344 amino acids and with the relative molecular weight as 38 570.53 Da and the theoretical isoe-lectric point as 9.59.The secondary structure and tertiary structure of CaWRKY39 were mainly composed of ir-regular curls,accounting for 60.76%.The phylogenetic analysis results indicated that CaWRKY39 had the clo-sest genetic relationship with cotton GhWRKY39,indicating that their function might be similar.Furthermore,in order to verify the target regulating function of CaWRKY39 to related genes,the CDS of CaWRKY39 gene was cloned by RT-PCR,and the yeast-one-hybrid vector pGADT7-CaWRKY39 was successfully constructed by In-Fusion cloning with NdeⅠ and Eco RⅠ for double digests.These results could provide the technical support and reference for further study on molecular mechanism of CaWRKY39 involved in the resistance to TSWV of pepper.
Pepper(Capsicum annuum L.)CaWRKY39Bioinformatics analysisYeast-one-hybrid pGADT7 vector construction
万方数据
维普
