Gene knock-in recombinant vector and preparation method thereof as well as method for preparing mouse model

A recombinant vector and gene knock-in technology, applied in the biological field, can solve the problems of inconvenient and flexible operation, high requirements for experimental instruments and equipment, and low fluorescence sensitivity, and achieve the effects of clean background, accurate experimental results, and high detection sensitivity
CN103642828AActive Publication Date: 2014-03-19BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
BIOCYTOGEN PHARMACEUTICALS (BEIJING) CO LTD
Publication Date
2014-03-19

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Abstract

The invention discloses a recombinant vector for gene knock-in. The recombinant vector comprises a DNA (deoxyribonucleic acid) fragment structure IL17A-IRES-luc, wherein IL17A is an encoding gene of interleukin 17A, IRES is internal ribosome entry site, and luc is an encoding gene of secreting luciferase. The invention also provides a method for preparing the gene knock-in recombinant vector and a mouse model prepared by using the recombinant vector, wherein the mouse model can be used for detecting the expression of the IL17A gene by detecting the expression of the secreting luciferase in blood or urine; a fluorescent microscope or other complicated equipment is not needed; the secreting luciferase emits light by virtue of an oxidizing reaction of substrate coelenterazine in the catalysis of luciferase, and laser light is not needed, so that the fluorescent background is far lower than GFP (green fluorescent protein) fluorescence, the detection sensitivity is higher, the background is cleaner, and the experiment results are more accurate.
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Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a method for preparing a mouse model and a gene knock-in recombinant vector. Background technique

[0002] Gene knock-in (Knockin) is a technology that introduces specific mutations or exogenous genes at the position of the target gene according to experimental requirements, such as introducing point mutations on the target gene (simulating a human genetic disease model); or introducing a reporter gene (such as EGFP, mRFP , mCherry, etc.) introduce specific sites of the target gene through homologous recombination, so that the expression of the target gene can be tracked and the expression profile of the gene can be studied through the expression of the reporter gene.

[0003] IL17A (Interleukin-17A) is a member of the IL-17 family of cytokines and plays a key role in host defense and inflammatory processes.

[0004] The current model mouse for monitoring IL17A gene expression is th...

Claims

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