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Functional Analysis And Protein Interaction Screening Of A Vacuole Sortung Protein AtVPS25in Aranbidopsis Thaliana

Posted on:2015-01-25Degree:MasterType:Thesis
Country:ChinaCandidate:M M GuoFull Text:PDF
GTID:2180330434960236Subject:Crop Genetics and Breeding
Abstract/Summary:PDF Full Text Request
Vps25constitutes an important constituent of the ESCRT complexes. There are fewresearches in function on Vps25in plants, therefore, studies on the function and proteininteraction screening of Arabidopsis AtVPS25analysis will help to explore the function ofVps25in plant, it has a certain theoretical significance from a number of biological processesin plant, such as autophagy and auxin transport. This study used Arabidopsis homozygousmutant vps25as material, used the methods of Arabidopsis vps25homozygous mutantphenotype identification and molecular biology for AtVPS25functional analysis, and yeasttwo-hybrid method for screening the interaction protein of AtVPS25, the main results are asfollows:1. Phenotype identification of vps25homozygous mutant in Arabidopsis found that,vps25represent inhibition in primary root elongation under stress conditions of auxin, nosignificant difference in the number of lateral roots; in the treatment conditions of carbon-limiting, Clo-0survival rate was40%, vps25mutant survival was5%, the chlorophyll contentin vps25was decreased significantly (P <0.01).2. Real-time PCR showed that under auxin stress conditions, in Clo-0, AtVPS25andAtAIR12were both induced in auxin response progress, and reached the highest expressionlevel at12h; in vps25, the AtAIR12expression was inhibited, AtVPS25has a positiveregulatory in expression of AtAIR12. In vps25, the expression level of auxin efflux carriergene AtPIN2was reduced and the expression level of auxin input carrier gene AtPIN2thegrowth hormone input carrier gene was ascendant.2. Making AtVPS25as bait protein, used the split-ubiquitin system screened theArabidopsis (Clo-0) genome-wide cDNA library, got78interacting proteins of AtVPS25.BiFC results showed that AtVPS25interacted with AtAIR12, and the interaction sites was onthe cell membrane and in the cytoplasm.
Keywords/Search Tags:Arabidopsis Thaliana, Vacuolar Sorting Protein, Functional Analysis, Protein Interaction
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