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Method for quantitative detection of target gene expression by flow cytometer

A technology of flow cytometry and gene expression, applied in the field of molecular biology, can solve problems that do not have the general applicability of other phenotype studies

Inactive Publication Date: 2017-07-25
DONGHUA UNIV
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Problems solved by technology

In this method, in addition to being quantified markers, fluorescent dyes also have the function of substrates, which are directly related to the functions being studied, so they do not have general applicability to other phenotypic studies

Method used

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  • Method for quantitative detection of target gene expression by flow cytometer
  • Method for quantitative detection of target gene expression by flow cytometer
  • Method for quantitative detection of target gene expression by flow cytometer

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Embodiment Construction

[0026] Below in conjunction with specific embodiment, further illustrate the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

[0027] A method for quantitative detection of target gene expression using flow cytometry, specifically as follows:

[0028]In cells expressing the target gene to be studied, use Madisen et al. (Madisen L, Zwingman TA, Sunkin SM, et al (2010). A robust and high-throughput Cre reporting and characterization system for the whole mouse brain, 13(1 ): 133-40.) The constructed Ai9-dtomato plasmid knocks in the red fluorescent repor...

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Abstract

The invention provides a method for quantitative detection of target gene expression by a flow cytometer. The method comprises knocking a red fluorescent protein gene into a cell expressing a target gene to be researched, knocking a green fluorescent protein gene into a target gene reading frame, synchronously expressing the red fluorescent protein gene, the green fluorescent protein gene and the target gene so that corresponding proteins are independently formed, screening external factors influencing the expression of the target gene through the cell synchronously expressing the red fluorescent protein gene and the green fluorescent protein gene, quantitatively detecting the intensity of a green fluorescent signal of the cell through a flow cytometer, carrying out homogenization through a red fluorescent signal, and quantitatively comparing the influence caused by the external factors on the target gene expression so that the external factors influencing the expression of the target gene are ensured. The method can compare expression amount difference of the specific gene under multiple treatment conditions and realizes high sensitivity screening of factors influencing the expression of the gene in high throughput and short time.

Description

technical field [0001] The invention relates to a method for quantitative detection of target gene expression by using a flow cytometer, and belongs to the technical field of molecular biology. Background technique [0002] Flow cytometry is a technique for one-by-one, multi-parameter, rapid qualitative and quantitative analysis or sorting of single cells or biological particles in a fast linear flow state. It has the advantages of fast detection speed, many measurement parameters, and a large amount of collected data , comprehensive analysis, high sorting purity, and flexible methods. Flow cytometry (FCM) is based on flow cytometry as the core technology, integrating physics, chemistry, biology, and multidisciplinary theories such as lasers and computers. The technical equipment integrated with technology is widely used in medical testing, cell biology, drug screening and other multidisciplinary fields. With the advancement of technology and the popularization of instrument...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6486
Inventor 周宇荀李晓宁王斯佳肖君华李凯
Owner DONGHUA UNIV
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