Screening and Identifying of Interaction Protein AtLBD15 in Arabidopsis thaliana

doi:10.7668/hbnxb.201751326

Abstract

Abstract: In order to further explore the role of LBD genes in Arabidopsis thaliana in response to plant abiotic stresses, we first inserted LBD15 gene into pGBKT7 vector and constructed the pGBKT7-LBD15 yeast expression vector. The pGBKT7-LBD15 was used to screen the interacted proteins of LBD15. The yeast two-hybrid technique was used to screen a full-length homogenized yeast library of Arabidopsis thaliana. There were 107 detected sequences in 146 samples of which 96 were identified as the Arabidopsis thaliana genes by NCBI Blast, accounting for 87.9% of the total. According to the gene accession number, 48 proteins were obtained, and 59% of them were duplicated. And then the protein sequences were searched via the website TAIR (www.arabidopsis.org/index.jsp) based on the gene accession number. The subcellular localization was analyzed on http://cello.life.nctu.edu.tw/. Through localization analysis, these proteins were mainly located in the chloroplast, cytoplasm and nucleus. Gene Ontology annotations using Blast2GO showed that the interacting proteins were involved in 14 biological processes, including cellular processes, metabolic processes, stimuli response, regulation of biological processes, single-tissue and multi-tissue processes, and developmental process regulation. Five of the possible interaction proteins were randomly selected and used to verify the interaction. The results showed that the colonies of AP19 protein and other proteins could grow normally on auxotrophic plates, further confirming that they were interacted proteins. The successful screening of yeast two-hybrid library laid the foundation for further study of the molecular mechanism of Arabidopsis LBD15.

Key words: Arabidopsis thaliana, LBD15, Yeast two-hybrid system, Protein interaction

CLC Number:

S54.03

GUO Zhaolai, SUN Xudong, YANG Yongping, XU Huini. Screening and Identifying of Interaction Protein AtLBD15 in Arabidopsis thaliana[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(5): 52-58. doi: 10.7668/hbnxb.201751326.

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