Cloning And Expression Analysis Of TaMYC1 Gene In Taraxacum Antungense Kitag.

MYC transcription factor is involved in the biosynthesis of plant secondary metabolism,however,the study of MYC transcription factor in Taraxacum aungue Kitag.has not been reported.Therefore,a MYC transcription factor was cloned from Taraxacum aungue Kitag.in this experiment,and did some research on it.the results are as follows:1.The Ta MYC1 gene was cloned from Taraxacum aungue Kitag.and its ORF was 1566 bp,which encoded 521 amino acids.The amino acid sequence translated by Ta MYC1 gene contains two typical conserved domains of basic-helix-loop helix(b HLH)and helix-loop-helix(HLH)of b HLH transcription factor family;further construction of phylogenetic tree found Ta MYC1 and GMYC1 gene in Gerbera jamesonii Bolus has the highest homology.The results of tissue-specific analysis showed that the relative expression of Ta MYC1 gene was highest in leaves.2.The prokaryotic expression vector p GEX4T-1-Ta MYC1 was constructed.After extraction and purification of the induced protein.The results showed that the purified GST-Ta MYC1 fusion protein was successfully obtained,and its size was about 85 k Da.3.The results of q RT-PCR showed that the relative expression of Ta MYC1 gene increased to 4.3 at 1 h and decreased to 4.7 at 3 h after treatment with Me JA.After treatment with Na Cl,the relative expression of Ta MYC1 gene increased to 19.3 at 1 h and increased to 42.7 at 3 h.After treatment with SA,the relative expression of Ta MYC1 gene increased to 5.3 at 1 h and it reached 6.7 at 3 h.This result indicates that the Ta MYC1 transcription factor is involved in the Me JA response pathway and also responds to SA hormone and salt stress signals.4.The overexpression vector p RI101-Ta MYC1 was constructed,and the genetic transformation was carried out by Agrobacterium-mediated method.Finally,the transgenic callus was obtained.The q RT-PCR results showed that the relative expression of Ta MYC1 gene in transgenic callus was 4.7.5.The promoter of MYC was cloned and the length was 679 bp;it was analyzed to determine that the promoter had cis-acting element elements such as MYB binding site and G-box.The p GADT7-Ta MYC1 and p Ab Ai-pro Ta MYC1 vectors were constructed,and yeast one hybridization confirmed the binding of Ta MYC1 to p Ab Ai-pro Ta MYC1.