Construction of Yeast Library and Under Salt Stress in Lilium pumilum Screening of LpNAC14 Interacting Proteins
This study aimed to investigate the signaling pathway of LpNAC14,a NAC transcription factor in Lilium pumilum,in response to salt stress,to analyze the protein interaction,so as to provide a basis for the further study of the functions of the related genes.RNA collected from the leaves and roots of L.pumilum under salt stress was used to construct a yeast two-hybrid cDNA library using the Gateway method.Two LpNAC14 gene fragments without self-activating activity were used to construct bait vec-tors,and then they were co-transformed into the library plasmids to screen for interaction proteins.The screened results were subjected to yeast validation and GO enrichment analysis,and one of the transcription factors,LpDi19-2,was selected for BiFC validation with LpNAC14.The yeast library was constructed with a library capacity of 1.12×107 CFU,with a recombination rate of 100%,and the average length of the in-serted fragments was greater than 1 000 bp.Nineteen proteins potentially interacting with LpNAC14 were preliminarily screened by library transformation.BiFC verification test showed that LpDi19-2 interacted with LpNAC14 in vivo.It is concluded that the quality of the constructed yeast library of L.pumilum un-der salt stress is high,which can meet the screening criteria,so that the screening results are reliable.This study provides a new way to explore the mechanism of LpNAC14 in response to salt stress.
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