Method for detecting promoter activity by utilizing double luciferase reporter genes
A dual-luciferase and reporter gene technology is applied in the field of using dual-luciferase reporter genes to determine the core promoter of Guanling cattle MSTN gene, which can solve the problems of complex interactions between regulatory elements and trans-acting factors, and achieve detection speed Fast, low-cost, high-sensitivity results
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[0021] The present invention can be better understood from the following examples. However, those skilled in the art can easily understand that the description of the embodiments is only for illustrating the present invention, and should not and will not limit the present invention described in the claims.
[0022] Embodiments of the present invention: a method for detecting promoter activity using a dual-luciferase reporter gene, specifically as follows:
[0023] 1. Preparation of strains, plasmids and reagents.
[0024] Strain: Escherichia coli competent cell DH5α was purchased from Dalian Takara Company;
[0025] Cell lines: mouse C2C12 cells and 3T3-L1 cells were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences;
[0026] Plasmids: Reporter vectors pGL3-Basic and pRL-TK were purchased from Promega.
[0027] Reagent source: 2×PCR Taq Mix was purchased from Tiangen Company;
[0028] T4 DNA Ligase and endotoxin-free plasmid mini-extraction kit were ...
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