Screening and Identification of the Interaction Proteins of the Transcriptional Regulator StFlo8 in Setosphaeria turcica
SUN He-He, WANG Cheng-Ze, JIA Jing-Zhe, SHEN Shen, HAO Zhi-Min*, DONG Jin-Gao*
State Key Laboratory of North China Crop Improvement and Regulation/Hebei Bioinformatic Utilization and Technological Innovation Center for Agricultural Microbes/Hebei Key Laboratory of Plant Physiology and Molecular Pathology, Hebei Agricultural University, Baoding 071000, China
Abstract：Flo8 (flocculation 8) gene, a key transcription regulatory factor downstream of the cAMP pathway, plays an important role in the regulation of fungal growth and development. In previous study, the involvement of StFlo8 in regulating the appressorium development and the formation of infection filaments in the early stage of Setosphaeria turcica was identified. In the present study, StFlo8 fused with glutathione S-transferase (GST) tag were heterologously expressed via the Escherichia coli expression system. Further, the GST pull down technology was used to screen the proteins that may interact with StFlo8 from the mycelial proteins of S. turcica, and the yeast two hybrid technology was used to verify the interaction of some of the proteins. The results showed that the target product with a size of 116 kD obtained by prokaryotic expression was consistent with the size of GST-StFlo8, 41 proteins that may interact with StFlo8 were obtained from the successfully purified expression product through GST pull down. After validating a subset of proteins using the yeast two-hybrid technique, it was revealed that Flo8 interacts with FAD-dependent oxidoreductase (GenBank No.165285). This study preliminarily clarified the interaction protein of StFlo8, providing informative references for further elucidating the regulatory molecular mechanism of the activity of StFlo8 in S. turcica.
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