Preparation method and application of an insect in vitro protein interaction detection system

A detection system and protein technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of high background, low efficiency of mammalian cell BIFC plasmid transfection screening and detection, complicated mammalian cell culture process, etc. To achieve the effect of effective technical means, low cost and simple method
CN104991072BInactive Publication Date: 2017-03-29NORTHWEST A & F UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
NORTHWEST A & F UNIV
Publication Date
2017-03-29
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a manufacturing method and application of an insect in-vitro protein interaction detecting system. At the positions of amino acid residues at the 159th bit and the 160th bit, mCherry is divided into a segment NmC and a segment CmC through a PCR method, and then the segment NmC and the segment CmC are each fused with a connecting zone sequence, a c-Myc label and a poly(A) sequence to form four fused segments; the four fused segments are each connected with a pIE-MCS plasmid with an autographa californica multiple nuclear polyhedrosis virus ie1 gene promoter and a gp64 gene poly(A) sequence to establish expression plasmids; the expression plasmids are combined to form an insect cell dimolecular fluorescent complementary detecting system. The method is simple and easy to implement, and the established dimolecular fluorescent complementary detecting system expands the application of an insect cell expression system and provides a convenient and effective technological means for insect proteomics and research on protein interaction between insects and pathogenic microorganisms of the insects.
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Description

technical field

[0001] The invention belongs to the technical field of bioengineering, and relates to a preparation method and application of an insect in vitro protein interaction detection system. Background technique

[0002] Under the normal physiological metabolic conditions of cells, proteins are the executors of cell activity and function. Through complex interactions, proteins participate in various life processes such as cell signal transduction, proliferation, differentiation, immunity, and apoptosis. In recent years, with the acquisition of genome and transcriptome information of a large number of species, proteomics—the systematic study of the protein structure, function, and interaction network of a species has become a hot spot in the field of life sciences. Among them, analyzing protein interactions is the basis for revealing protein structure and function, which helps to systematically explain various life phenomena and processes, explore the occurrence and ...

Claims

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