Kit and method for detecting expression level of TUBB3 (Tubulin B3) mRNA (messenger Ribonucleic Acid)

A technology of gene expression and kit, which is applied in the field of kits for detecting TUBB3 mRNA gene expression, can solve the problem of poor curative effect of anti-microtubule chemotherapy

Inactive Publication Date: 2013-05-08
周宏灏
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  • Abstract
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  • Claims
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Problems solved by technology

However, patients with high expression of TUBB3 have poor efficacy of anti-microtubule chemotherapy (Pascal S and Charles D. Lancet Oncol. 2008;9:168-75.)

Method used

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  • Kit and method for detecting expression level of TUBB3 (Tubulin B3) mRNA (messenger Ribonucleic Acid)
  • Kit and method for detecting expression level of TUBB3 (Tubulin B3) mRNA (messenger Ribonucleic Acid)
  • Kit and method for detecting expression level of TUBB3 (Tubulin B3) mRNA (messenger Ribonucleic Acid)

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Embodiment Construction

[0026] In order to make the object, technical solution and advantages of the present invention clearer, preferred embodiments of the present invention are described in detail below.

[0027] 1. Configuration of fluorescent quantitative PCR detection kit for TUBB3 gene

[0028] The fluorescent quantitative PCR detection kit of the TUBB3 gene of the present embodiment comprises the following components:

[0029] 1) TUBB3 gene standard product: recombinant pUC57 vector containing the TUBB3 gene coding sequence (GeneBank number NM_001197181.1 from the 339th to the 436th base);

[0030] 2) Detect the upstream and downstream primers of TUBB3 gene:

[0031] TUBB3-upstream primer: 5'-GGCCTCTTCTCCACAAGTACG-3' (SEQ ID NO.2)

[0032] TUBB3-downstream primer: 5'-GAAGAGATGTCCAAAGGCCC -3' (SEQ ID NO.3);

[0033] 3) 5× reverse transcription buffer, reverse transcription primer Oligo-(dT)16 at a concentration of 10 pmol / μl, dNTPs solution at a concentration of 20 pmol / μl, M-MLV reverse tra...

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Abstract

The invention discloses a kit and method for detecting the expression level of TUBB3 (Tubulin B3) mRNA (messenger Ribonucleic Acid). The kit comprises a standard substance and an internal reference gene real-time quantitative PCR (Polymerase Chain Reaction) system, wherein the standard substance is used for making a standard curve. Through the method, the expression level of TUBB3 can be accurately, rapidly and quantitatively determined. If the method and the kit, disclosed by the invention, are applied in clinic, doctors can rationally select anti-TUBB3 for treatment.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit and method for detecting TUBB3 mRNA gene expression. Background technique [0002] Anti-microtubule drugs are a class of drugs that target cellular tubulin. A class of broad-spectrum chemotherapeutic drugs that affect the formation of spindles and inhibit cell mitosis by acting on cellular microtubules. Currently commonly used anti-microtubule drugs mainly include paclitaxel, docetaxel, vinblastine, vincristine, and vinorelbine. After paclitaxel / docetaxel acts on tumor cells, it can promote the polymerization of microtubules in tumor cells and stabilize the polymerized microtubules, resulting in the accumulation of a large number of microtubules in the cells, thereby interfering with various functions of cells, especially stopping cell division . Vinblastine / vincristine can inhibit the polymerization of tubulin in tumor cells, inhibit the formation of spindle mic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 张伟郭成贤陈小平刘昭前周宏灏
Owner 周宏灏
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