Construction of cDNA library of Bipolaris sorokiniana and screening of BsTup1 interacting proteins
Journal Title: Journal of Henan Agricultural University - Year 2024, Vol 58, Issue 2
Abstract
[Objective] To investigate the molecular mechanisms of growth, development, and pathogenicity of Bipolaris sorokiniana and identify the interacting proteins of BsTup1. [Method] We used spores and mycelia at different stages of B. sorokiniana as materials to construct a yeast two-hybrid cDNA library, and screened the yeast two-hybrid library by BsTup1 gene as the bait for determining the proteins interacting with BsTup1. [Result] 1) The mixed full-length cDNA library of conidia and mycelium was successfully constructed for the first time using SMART (switching mechanism at 5′ end of the RNA transcript) technology. The results of library identification showed that the constructed cDNA library had a capacity of 4.8×107 cfu·mL-1, the library insertion fragment recombination rate reached 100% and the average size was around 1 000 bp. 2) The bait protein vector pGBKT7-BsTup1, a transcriptional inhibitor of BsTup1 in B. sorokiniana, successfully constructed and confirmed no self-activation activity. 3) Using this bait vector, 38 candidate proteins interacting with BsTup1 were obtained after sequencing, sequence alignment, and yeast verification. [Conclusion] We successfully constructed a cDNA library of B. sorokiniana, and identified 38 candidate proteins which interacted with BsTup1.
Authors and Affiliations
Changshui LI, Yuehua GENG, Meng YAO, Bingsen ZHAO, Shunpei XIE, Chao XU, Qingzhou MA, Meng ZHANG
Keywords
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- EP ID EP769332
- DOI 10.16445/j.cnki.1000-2340.20240030.001
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