Dual Expression Vector for High-Throughput cDNA Interaction Screening

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Solution Overview

Problem

Traditional yeast two-hybrid systems require constructing multiple vectors for testing multiple protein interactions, significantly increasing workload and limiting efficiency.

Innovation Solution

A dual expression vector is used to simultaneously express two cDNA libraries, allowing for the direct testing of protein interactions by transforming the vector into yeast and observing growth on selective culture media.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If multiple vectors are constructed for testing multiple protein interactions, then the coverage of protein interaction testing is improved, but the workload and complexity of vector construction increases significantly

Engineering Contradiction:

Improvecoverage of protein interaction testingVSAvoidcomplexity of vector construction

Core Design Contradiction:

Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent combines multiple vector functions into a single dual expression vector that can simultaneously accommodate both bait and prey proteins. This single vector system replaces the traditional requirement for multiple separate vectors, thereby maintaining comprehensive protein interaction testing capability while significantly reducing vector construction complexity and workload.

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The dual expression vector is designed with multi-functionality to serve multiple purposes: it can express both bait and prey proteins simultaneously, work with multiple cDNA libraries, and enable various types of protein interaction screenings. This universal vector design eliminates the need to construct different vectors for different testing scenarios.

Inventive Principle:

Principle #6Universality (Multi-functionality)

2Adaptability or versatility

If multiple vectors are constructed for testing multiple protein interactions, then the coverage of protein interaction testing is improved, but the time required for identification increases

Engineering Contradiction:

Improvecoverage of protein interaction testingVSAvoidtime required for identification

Core Design Contradiction:

Adaptability or versatilityVSLoss of time

Solution Approach 1:

By merging the functionality of multiple vectors into a single dual expression vector, the patent enables simultaneous expression and screening of multiple protein interactions in one experimental system. This eliminates the sequential time required for constructing and testing multiple separate vectors, thereby maintaining comprehensive testing coverage while significantly reducing the total identification time.

Inventive Principle:

Principle #5Merging (Combining)

3Reliability

If traditional yeast two-hybrid system is used for library interactions, then the ability to detect protein interactions is maintained, but the efficiency of identification is severely limited

Engineering Contradiction:

Improveability to detect protein interactionsVSAvoidefficiency of identification

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The dual expression vector maintains the reliable protein interaction detection capability of traditional yeast two-hybrid systems while adding multi-functionality to handle library-scale screenings. The vector preserves the GAL4-based detection mechanism for reliability while enabling high-throughput application with multiple cDNA libraries, thereby significantly improving identification efficiency without sacrificing detection accuracy.

Inventive Principle:

Principle #6Universality (Multi-functionality)

Data Source

PatentUS20260109966A1METHODS FOR PREPARING INTERACTION PLASMID LIBRARY AND TESTING INTERACTION BETWEEN cDNA LIBRARIES

Publication Date: 2026.04.23 HUAZHONG AGRI UNIV

AI summary

A method for preparing interaction plasmid library and a method for testing interactions between cDNA libraries are provided. The cDNA libraries could be inserted into the first multiple cloning site and the second multiple cloning site of the dual expression vector for testing the interactions between the different cDNA libraries.