Construction of Yeast Two-hybrid Bait Vector and Identification of Proteins Interacting with <i>BsMYB62</i> in <i>Begonia semperflorens</i>

LIU Jing; WANG Yang; YAO Kexin; QU Ying; WANG Ruibo; LIU Shengnan; LI Yonghua; ZHANG Kaiming

doi:10.19303/j.issn.1008-0384.2021.04.006

Volume 36 Issue 4

Apr. 2021

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Article Navigation > Fujian Journal of Agricultural Sciences > 2021 > 36(4): 418-425

LIU J, WANG Y, YAO K X, et al. Construction of Yeast Two-hybrid Bait Vector and Identification of Proteins Interacting with BsMYB62 in Begonia semperflorens [J]. Fujian Journal of Agricultural Sciences，2021，36（4）：418−425 doi: 10.19303/j.issn.1008-0384.2021.04.006

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LIU J, WANG Y, YAO K X, et al. Construction of Yeast Two-hybrid Bait Vector and Identification of Proteins Interacting with BsMYB62 in Begonia semperflorens [J]. Fujian Journal of Agricultural Sciences，2021，36（4）：418−425 doi: 10.19303/j.issn.1008-0384.2021.04.006

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Construction of Yeast Two-hybrid Bait Vector and Identification of Proteins Interacting with BsMYB62 in Begonia semperflorens

doi: 10.19303/j.issn.1008-0384.2021.04.006

College of Landscape Architecture and Arts, Henan Agriculture University, Zhengzhou, Henan　450002, China

Received Date: 2020-09-04
Rev Recd Date: 2020-12-18

Available Online: 2021-03-27

Publish Date: 2021-04-30

Abstract

Abstract

Objective The selected transcriptomes of Begonia semperflorens that can trigger the anthocyanin biosynthesis in the plant was used to clone the MYB transcription factor, and the cDNA databank applied to identify the potential interacting proteins associated with the biological process. Method Transcriptomes of B. semperflorens plant exposed to low temperature or intense light to induce the anthocyanin biosynthesis that turns the leaves red were screened for positive identification and cloning the full length cDNA of an MYB transcription factor, BsMYB62 (Gene ID: MT560845). Bioinformatics and potential interacting proteins of BsMYB62 were obtained with the designed primers and cloned cDNA. A pGBKT7-BsMYB62 bait vector was constructed and tested for toxicity and auto-activation. From the yeast two-hybrid library using the mating method, potential interacting proteins were named. Result The full length cDNA of BsMYB62 was determined to be 801 bp. The pGBKT7-BsMYB62 was found not toxic to the host yeast and capable of significantly inhibiting auto-activating in SD/-Trp-Leu-His-Ade medium including 45 mM of 3-AT. Nine potential interacting proteins were identified to include XTH9, LHB1B2, EBS7, PSI-F, UBE2, FDH, RBCS1A, FP6, and BCA. Conclusion It was speculated that BsMYB62 of B. semperflorens interacted with 9 proteins in response to the stress of low temperature or intense light.
- Begonia semperflorens,
- BsMYB62,
- yeast two-hybrid,
- interaction protein

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References(25)

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