Efficient yeast two-hybrid three-frame library construction technology

A yeast two-hybrid and library construction technology, which is applied in the field of efficient yeast two-hybrid three-frame library construction, can solve the problem of high false negatives in the library, achieve the effects of reducing false negative rates, avoiding mutations, and ensuring sample uniformity

Inactive Publication Date: 2021-03-30
WUHAN GENECREATE BIOLOGICAL ENG CO LTD
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Problems solved by technology

The above-mentioned library has the problem of high false negative

Method used

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  • Efficient yeast two-hybrid three-frame library construction technology
  • Efficient yeast two-hybrid three-frame library construction technology
  • Efficient yeast two-hybrid three-frame library construction technology

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Embodiment Construction

[0027] The principles and features of the present invention are described below, and the examples are only used to explain the present invention, but not to limit the scope of the present invention.

[0028] The present invention designs a recombinant vector for the construction of a high-efficiency yeast two-hybrid triple-box library. There are three types of recombinant vectors. The three types of recombinant vectors are inserted into the original PGADT7 vector with base sequence 1, base sequence 2 and nucleotide sequence 2 respectively. The base sequence 3 is constructed, the nucleotide sequence of the base sequence 1 is SEQ ID NO: 1, the nucleotide sequence of the base sequence 2 is SEQ ID NO: 2, and the base sequence 3 The nucleotide sequence is SEQ ID NO:3.

[0029] At present, the key to limit the application of yeast two-hybrid technology lies in the quality of the library, and the existing yeast two-hybrid libraries all have the problem of false negatives. The invent...

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Abstract

The invention relates to an efficient yeast two-hybrid three-frame library construction technology which comprises three recombinant vectors for constructing a high-efficiency yeast two-hybrid three-frame library, and the three recombinant vectors are constructed by respectively inserting a base sequence 1, a base sequence 2 and a base sequence 3 into an original PGADT7 vector. The nucleotide sequence of the base sequence 1 is SEQ ID NO: 1, the nucleotide sequence of the base sequence 2 is SEQ ID NO: 2, and the nucleotide sequence of the base sequence 3 is SEQ ID NO: 3. The invention also discloses a construction method of the vector and a construction method for constructing the yeast two-hybrid three-frame library by using the vector. The yeast two-hybrid three-frame library is constructed through the recombinant vector, the false negative rate can be effectively reduced, and the cost is low.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an efficient yeast two-hybrid triple-frame library construction technology. Background technique [0002] Yeast two-hybrid technology has a wide range of applications in studying the interaction between two known proteins and using yeast two-hybrid technology for high-throughput screening of known unknown proteins, and has made great progress in recent years. The optimization and improvement of yeast two-hybrid technology has been designed to reduce the production of false positives or false negatives in yeast two-hybrid screening. The quality of the library is the key to achieve the above goals. [0003] In the mat&plate library developed by Clontech, the yeast prey protein library and the bait strain are co-cultured on a suitable selection medium to screen out prey proteins that interact with known proteins. The principle of its library construction is to utilize the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/81C12N15/66C12N15/10C40B50/06
CPCC12N15/81C12N15/1096C40B50/06C12Q2531/113
Inventor 李昆鹏李敏
Owner WUHAN GENECREATE BIOLOGICAL ENG CO LTD
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