Screening of Potato (Solanum tuberosum) Proteins interacting with StLURP1 Gene by Yeast (Saccharomyces cerevisiae) Two-Hybrid System
FANG Chen-Xi1, CHE Yan1, LIAO Yu-Qiu1, WANG Yi-Fan1, ZHANG Ning1,2, SI Huai-Jun1,2,*
1 College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China; 2 Gansu Provincial Key Laboratory of Aridland Crop Science, Lanzhou 730070, China
Abstract：The late up-regulated in response to Hyaloperonospora parasitica gene 1(LURP1) plays a key role in the response against pathogenic oomycete H. parasitica and resistance to stress in plants. In order to elucidate the biological function of the potato (Solanum tuberosum) LURP1 gene and to clarify its position in the protein interaction network, the bait vector pGBKT7-StLURP1 was constructed by homologous recombination and used to screen the potato cDNA library via the yeast (Saccharomyces cerevisiae) two-hybrid (Y2H) system in this research. The result showed that the positive rate of Y2H library screening was about 70%, and finally 88 positive clones were yielded, which was corresponded to 12 StLURP1 interacting proteins with complete ORF. The authenticity of the interactions was verified by retest. Blast alignment and Gene ontology (GO) annotation analysis demonstrated that StLURP1 could interact with the proteins such as DnaJ family protein, oxidoreductase/transcriptional repressor, glycerophosphodiester phosphodiesterase (GPGD), polyphenoloxidase, proline-rich protein, heavy metal transport/detoxification domain-containing protein, actin depolymerizing factor (ADF), nonspecific serine/threonine or tyrosine protein kinase, et al. The result indicated that StLURP1 may be involved in multiple biological processes, including lipid metabolism, biotic and abiotic stress responses, actin depolymerizing, heavy metal ions transport, protein folding and other biological processes. The present study would provide a theoretical foundation for further studying of biological function and role of StLURP1 gene.
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