Nucleic acid molecule, corresponding vector, cell for stable co-expression of GFP-Flag-FXR and preparation method

Abstract

The invention provides a nucleic acid molecule, a corresponding carrier, a cell for stable co-expression of GFP-Flag-FXR and a preparation method. The nucleic acid molecule comprises an FXR sequence for expressing FXR protein and / or a corresponding complementary sequence, a Flag sequence for expressing a Flag tag and / or a corresponding complementary sequence, and a GFP sequence for expressing a GFP tag and / or a corresponding complementary sequence. The amino acid sequence of the FXR protein is shown as SEQ ID NO: 1, the amino acid sequence of the Flag tag is shown as SEQ ID NO: 2, and the amino acid sequence of the GFP tag is shown as SEQ ID NO: 3. The nucleic acid molecule can simultaneously express a GFP tag, a Flag tag and an FXR protein. GFP is used as a label, the transfection efficiency of FXR can be observed in real time in the transfection process, and the operation efficiency is improved; on the premise that the basic structure of the FXR is not changed, the Flag tag of the small molecule can be used for basic researches such as IP, ChIP and the like which have high requirements on protein molecule conformation, and the research range can be expanded.

Description

technical field [0001] The invention relates to the technical field of cell engineering, in particular to a nucleic acid molecule, a corresponding carrier, a cell stably co-expressing GFP-Flag-FXR and a preparation method. Background technique [0002] Obesity is a common and frequently-occurring disease worldwide, which can easily lead to various complications such as cardiovascular disease, diabetes, and intestinal injury. Bile acids and their receptors play key roles in glucose and lipid metabolism, intestinal injury repair and other aspects in humans and animals. FXR is a key nuclear receptor for bile acids. FXR in the intestine, on the one hand, is responsible for regulating the expression of downstream cytokines such as FGF15 / 19, thereby regulating blood lipids; on the other hand, it may also be responsible for regulating the proliferation and differentiation of intestinal cells, thereby regulating the normal development of human and animal intestines. Because human ...

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Problems solved by technology

However, limited by factors such as cell source and species, the use of selective activators will cause problems such as effects, and at the same time, there is also the problem that it is impossible to study the downstream regulatory network of FXR

[0006] At present, the patent for FXR gene expression in zebrafish is undergoing substantive examination, but because the gene sequence of zebrafish is quite different from that of mammals, it is of limited significance in related research

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