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Drug screening method based on p16 promoters

A promoter, p16ink4a technology, used in biochemical equipment and methods, microbial assay/inspection, introduction of foreign genetic material using vectors, etc.

Inactive Publication Date: 2011-11-23
乐清市康诺生物医药公共服务平台有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

p16 INK4a As a node gene in the cell signaling network, a gene is abnormally expressed in many tumor cells, and the activity of its promoter directly affects the expression level of the gene, but there are few reports on anti-tumor drug screening using the promoter as a target

Method used

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  • Drug screening method based on p16 promoters
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  • Drug screening method based on p16 promoters

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Embodiment Construction

[0042] The embodiments of the present invention are described in detail below. This embodiment is implemented on the premise of the technical solution of the present invention, and detailed implementation methods and specific operating procedures are provided, but the protection scope of the present invention is not limited to the following implementation example.

[0043] 1.p16 INK4a Cloning and identification of promoters.

[0044] p16 INK4a Promoter / luciferase (luciferase) eukaryotic expression vector, is characterized in that: named p16 INK4a -luciferase eukaryotic expression vector, wherein p16 INK4a - The full length of the luciferase sequence is 3300bp, of which p16 is between 138 and 1478 INK4a Promoter, 1535-3187 is the luciferase reading frame.

[0045] p16 INK4a The construction method of promoter / luciferase (luciferase) eukaryotic expression vector is characterized in that it comprises: designing primers from human ovary surface epithelial cell genome (HOSE D...

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Abstract

The invention relates to a drug screening method based on p16 promoters, belonging to the technical field of drug preparation. The sequence of the p16<INK4a> promoter-luciferase eukaryon expression vector is disclosed as Seq ID NO.1; the full length of the sequence is 3300bp, wherein 138th-1478th base pairs are p16<INK4a> promoters, and 1535th-3187th base pairs are luciferase reading frames; and the eukaryon expression vector can be used for detecting the anticancer activity of a compound.

Description

technical field [0001] The present invention relates to a screening method in the technical field of drug preparation and its preparation and application, in particular to a p16-based method in osteosarcoma U2OS cell lines INK4a (Cyclin-dependent kinase CDK4 inhibitor) promoter luciferase (luciferase) anti-tumor drug screening method and its construction method and application. Background technique [0002] Cancer is a group of cells that deviate from the normal growth cycle, and many of these cells have altered genes. In the experiment, the tumor suppressor gene p16, which has significant changes in the occurrence of cancer and has a significant impact on the occurrence of cancer, is selected. INK4a . The promoter part is amplified, and an easy-to-observe signal protein gene—luciferase gene (luciferase) is connected downstream of it. After 24 hours of transient transfection of U2OS or other cancer cell lines, a certain concentration of the compound was added, and the eff...

Claims

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Application Information

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IPC IPC(8): C12N15/79C12N15/63C12N15/66C12Q1/66C12Q1/02G01N21/64
Inventor 李大伟蔺剑武正华施丽丹金龙
Owner 乐清市康诺生物医药公共服务平台有限公司
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