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Method for detecting transcription level of tree shrew xanthine dehydrogenase/oxidase gene through RT-PCR

A xanthine dehydrogenase and gene transcription technology, applied in the field of molecular biology, achieves the effects of simple method, strong primer specificity, and low detection cost

Active Publication Date: 2014-11-26
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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Problems solved by technology

At present, there is no semi-quantitative method for xanthine oxidase / dehydrogenase transcription level using tree shrews at home and abroad

Method used

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  • Method for detecting transcription level of tree shrew xanthine dehydrogenase/oxidase gene through RT-PCR
  • Method for detecting transcription level of tree shrew xanthine dehydrogenase/oxidase gene through RT-PCR
  • Method for detecting transcription level of tree shrew xanthine dehydrogenase/oxidase gene through RT-PCR

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Embodiment Construction

[0039] The experimental methods used in the following examples are conventional methods unless otherwise specified.

[0040] The materials, reagents and the like used in the following examples were obtained from commercial sources unless otherwise specified.

[0041] 1. Experimental animals used in this example:

[0042] Chinese and Burmese tree shrews of the Dianxi species are raised at a normal level. The temperature of the breeding environment is 20°C to 25°C and the humidity is 40% to 70%. They are fed normally and drink water freely. The weight is 110-150g, 1 to 3 years old, half male and half male, fed with conventional and unified steamed cake feed.

[0043] 2. Preparation of various solutions

[0044] Preparation of 1% sodium carboxymethylcellulose (abbreviated as 1% CMC-Na): Weigh 3g of CMC-Na (manufactured by Kunming Chengyue Technology Co., Ltd.), put it into a 500ml Erlenmeyer flask, slowly add 100ml of boiled Bacterial distilled water swells, then slowly add 1...

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Abstract

The invention provides a method for detecting the transcription level of tree shrew xanthine dehydrogenase / oxidase gene through RT-PCR. The method comprises the following steps: carrying out PCR amplification by using a PCR primer combination with cDNA obtained after inverse transcription of total RNA of a sample as a template to obtain a ratio of the gray value of an amplification product of an XDH / XO gene fragment to the gray value of a PCR amplification product of an internal control gene GAPDH, calculating the relative expression value of XDH / XO mRNA under different physiological conditions according to the gray value ratio, and semi-quantitatively detecting the transcription level of tree shrew xanthine dehydrogenase / oxidase gene according to the calculated relative expression value in order to provide an effective way to study the functions of tree shrew xanthine oxidase and the influences of related drugs on the tree shrew xanthine oxidase. The method is suitable for the semi-quantitative RT-PCR detection, and has the advantages of low cost, simple operation, high repeatability low detection cost, and good specificity and sensitivity.

Description

technical field [0001] The invention relates to a detection method, in particular to a method for detecting the transcription level of tree shrew xanthine dehydrogenase / oxidase by using a semi-quantitative RT-PCR method, and belongs to the field of molecular biology technology. Background technique [0002] Xanthine dehydrogenase / oxidase (xanthine dehydrogenase / oxidase, XDH / XO ) is a non-specific aerobic dehydrogenase, which catalyzes the last two steps of purine metabolism in the process of purine metabolism, and forms the final product of uric acid metabolite uric acid from xanthine and hypoxanthine, which plays an important role in the metabolic process of uric acid It is a very important enzyme in the metabolism of uric acid in the body. [0003] In recent years, with the improvement of people's living standards, the incidence of hyperuricemia has increased year by year, and it is often accompanied by hypertension, hyperlipidemia and diabetes. According to domestic and...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q2545/101
Inventor 唐东红叶尤松罕园园李桂珍杨光蕊贾学云李润平念昕
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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