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Recombinant plasmid containing HDAC1 gene promoter and reporter gene as well as construction and application of recombinant plasmid

A technology of recombinant plasmid and reporter gene, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial measurement/testing, etc., can solve the problems of endangering human life and health, cancer recurrence and metastasis, etc., and achieves high sensitivity and start-up The effect of reduced child activity and low cost

Pending Publication Date: 2021-10-22
XUZHOU MEDICAL UNIV
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Problems solved by technology

However, cancer is extremely prone to recurrence and metastasis, which still seriously endangers human life and health[2,3]

Method used

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  • Recombinant plasmid containing HDAC1 gene promoter and reporter gene as well as construction and application of recombinant plasmid
  • Recombinant plasmid containing HDAC1 gene promoter and reporter gene as well as construction and application of recombinant plasmid
  • Recombinant plasmid containing HDAC1 gene promoter and reporter gene as well as construction and application of recombinant plasmid

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Embodiment Construction

[0047] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0048] 1. Construction of recombinant plasmids

[0049] 1) Mass amplification and extraction of pGL3-basic plasmid:

[0050] The map of pGL3-basic plasmid is as follows figure 1 As shown, conventionally prepared DH5α competent cells were transformed with the pGL3-basic plasmid, and a large amount of amplification was performed. The amplified plasmid was verified by agarose gel electrophoresis, and the results were as follows figure 2 As shown; a large number of pGL3-basic plasmids were prepared by alkaline lysis, and the purity and content of the plasmids were detected by electrophoresis; the pl...

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Abstract

The invention discloses a recombinant plasmid containing an HDAC1 (histone deacetylase 1) gene promoter and a reporter gene as well as a construction method and the application of the recombinant plasmid. A promoter specific sequence of a histone deacetylase 1 gene is cloned and recombined with the promoter specific sequence to a firefly luciferase reporter gene vector so as to construct a recombinant plasmid vector of the HDAC1 reporter gene; the constructed vector and a reporter gene vector of renilla luciferase are co-transfected into human prostate cancer cells PC3, and the transcriptional activity of an active reaction HDAC1 gene promoter of firefly and renilla luciferase is detected at the same time, so that a method for targeted screening of tumor inhibiting drugs is established. The method has excellent application prospects in the aspects of screening tumor inhibiting drugs, evaluating the possible influence of to-be-detected drugs on tumors, researching the action mechanism of the drugs and the like.

Description

technical field [0001] The invention belongs to the field of antitumor drug screening in the field of biotechnology, and in particular relates to a recombinant plasmid containing an HDAC1 gene promoter and a reporter gene, a construction method and application thereof. Background technique [0002] At present, cancer has become the primary disease that endangers human life and health. The low early detection rate of cancer leads to unsatisfactory treatment effects and poor prognosis. With the continuous development of radiotherapy, chemotherapy, surgery and other treatment methods, the survival period of cancer patients has been prolonged. However, cancer is extremely prone to recurrence and metastasis, which still seriously endangers human life and health [2,3]. Finding a method that can effectively inhibit the occurrence or metastasis of cancer is undoubtedly the gospel of mankind. Therefore, it is extremely important to create a high-throughput anti-tumor drug screening ...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/113C12N15/65C12Q1/66
CPCC12N15/85C12N9/80C12N15/65C12Q1/66C12Y305/01098
Inventor 刘志伟蒋昕娈袁琪徐妍李梦伟朱立东
Owner XUZHOU MEDICAL UNIV
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