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| Identification of Transcription Activity Region and Screening of Interaction Proteins of SmMYC2 Transcription Factor in Salvia miltiorrhiza |
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Abstract Myelocytomatosis protein2 (MYC2) is the core transcription factor of jasmonic acid signaling pathway and involves in plant defense response, secondary metabolism regulation, growth and development. Screening interaction proteins of Salvia miltiorrhiza transcription factor SmMYC2 is helpful to further study the biological function of SmMYC2. The MYC2 gene in S. miltiorrhiza was cloned and constructed into pGBKT7-GW (BD) to obtain the yeast (Saccharomyces cerevisiae) bait expression vector of BD-SmMYC2. The cytotoxicity and self-activation of BD-SmMYC2 were detected. The results showed that SmMYC2 has strong self-activation activity. In order to analyze the self-activation region, the N-terminal region (including the bHLH-MYC-N superfamily domain) and the C-terminal region (including the HLH domain) of SmMYC2 were cloned and constructed into the yeast expression vector BD. The transcription activity domain of the SmMYC2 was analyzed through detecting the transcription activity of the N-terminus and C-terminus of the SmMYC2. The SmMYC2-N-BD yeast turned blue on SD/-Trp/X-α-gal medium and could grow on SD-Trp-His and SD-Trp-Ade mediums, However, the SmMYC2-C-BD yeast was colorless on SD/-Trp/X-α-gal medium and no growth on SD-Trp-His and SD-Trp-Ade medium. These results indicated that the N-terminus of SmMYC2 has obvious self-activation activity and the C-terminus of SmMYC2 had no self-activation activity. The interactive proteins were screened from cDNA library of S. miltiorrhiza using yeast two-hybrid system through the bait vector of SmMYC2-C-BD. Function annotation showed that these candidate interaction proteins included WRKY protein containing the WRKYGQK domain, the peroxiredoxin reductase of Prxs protein family and succinate semialdehyde dehydrogenase. The experimental results provide basic date for further studying on the interaction proteins of MYC2.
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Received: 12 March 2018
Published: 26 September 2018
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