A yess-based method for analyzing protein interactions in prokaryotes

A protein interaction, prokaryotic technology, applied in the field of protein interaction analysis in prokaryotes based on YESS, can solve problems such as false positives and false negatives, inappropriate proteins, and inability to quantify the strength of protein interaction relationships.
CN109872774BActive Publication Date: 2021-04-02武汉睿嘉康生物科技有限公司
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
武汉睿嘉康生物科技有限公司
Publication Date
2021-04-02

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Abstract

The invention belongs to the technical field of gene engineering, and discloses a method for analyzing protein interaction in prokaryotes based on a Yeast Endoplasmic reticulum Sequestration System (YESS). The method includes the steps: constructing an expression plasmid pESD-PPI-GFP-mCherry capable of performing Golden Gate assembling, with gfp and mCherry fluorescent genes, based on pESD-PPI plasmid; selecting 5 mutually interacted protein pairs from the existing data of Zymomonas mobilis on a UniProt database and the protein pairs which are not reported to have interaction as positive and negative contrasts for studying protein interactions of proteins in Zymomonas mobilis; and quantifying the analysis results. The method for analyzing protein interactions in prokaryotes can achieve high-throughput and quantitative study on protein interaction in Zymomonas mobilis. The method for analyzing protein interactions in prokaryotes based on YESS has many advantages of being simple, being easy to operate, being highly efficient, being sensitive and the like.
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Description

technical field

[0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a method for analyzing protein interaction in prokaryotes based on YESS. Background technique

[0002] At present, the existing technologies commonly used in the industry are as follows:

[0003] The study of protein-protein interactions is of great significance in many biological fields, and it is very important for the analysis of various metabolic processes, regulatory networks and molecular mechanisms of energy conversion in cells.

[0004] At present, methods for verifying protein-protein interactions mainly include several classic experimental techniques: yeast two-hybrid technique, E. coli hybridization technique, pull-down technique and so on. Yeast two-hybrid technology is characterized by sensitivity, high efficiency, and high-throughput automation. However, there are many shortcomings in the actual application process, such as too high false n...

Claims

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