Improved bimolecular fluorescence complementation carrier for researching protein interaction

Abstract

The invention provides an improved bimolecular fluorescence complementation carrier for researching the protein interaction. The carrier includes a CaMV 35S promoter and a CaMV 35S terminator, and a cEYFP box or a nEYEP box is included between the CaMV 35S promoter and the CaMV 35S terminator. The construction cost of the carrier is low, while the construction efficiency is high.

Description

technical field [0001] The invention relates to a carrier and a method for preparing the carrier, belonging to the technical field of genetic engineering. Background technique [0002] Protein interaction is a basic physiological function in cells, which occurs in every physiological process in all subcellular structures and organelles. Clarifying when and where protein interaction occurs and how protein complexes are formed can provide a basis for determining Protein biological roles provide decisive clues. [0003] Bimolecular fluorescence complementation (BiFC) is a technology that uses the luminescent principle of fluorescent proteins to study protein interactions. The main process is to cut fluorescent proteins into two parts, the N-terminal and the C-terminal, and link them to the target protein respectively. The recombinant vector co-transfects cells. If there is an interaction between the target proteins, the N-terminal fragment and the C-terminal fragment of the fl...

AI Technical Summary

Problems solved by technology

This method has some disadvantages: First, rare restriction endonucleases are relatively expensive, and it is not easy to digest and connect, which will greatly increase the difficulty in the research process

Secondly, it is connected to the intermediate vector and the binary expression vector successively. This process is relatively complicated and cumbersome.

Another method is to use the GATEWAY carrier. Although this method is fast and efficient, the reagents used are expensive

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