RT-qPCR detection method of abcg2 gene transcription level in macaques

A technology of gene transcription and rhesus monkey, which is applied in biochemical equipment and methods, microbial measurement/testing, etc., to achieve high specificity, simple method and high repeatability

Active Publication Date: 2022-04-15
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

At present, there is no use of rhesus monkeys at home and abroad. ABCG2 Research on Quantitative Detection Method of Transcript Level

Method used

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  • RT-qPCR detection method of abcg2 gene transcription level in macaques
  • RT-qPCR detection method of abcg2 gene transcription level in macaques
  • RT-qPCR detection method of abcg2 gene transcription level in macaques

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Embodiment

[0051] A RT-qPCR detection in macaques ABCG2 A method at the level of gene transcription, comprising the steps of:

[0052] (1) Design the following primers:

[0053] According to the macaca mulatta monkey in the NCBI gene bank ABCG2 Nucleotide sequence number: NM_001032919.1, rhesus monkey ( macaca mulatta )of GAPDH Nucleotide sequence number NM_001195426.1 designed primers, the above primers were synthesized by Shanghai Bioengineering Co., Ltd.:

[0054] Macaque ABCG2 The specific upstream and downstream primers for gene expression level are:

[0055] ABCG2 F: 5’-TTAGCTGCAAGGAAAGATCCAAG-3’

[0056] ABCG2 R: 5'-GTCATAGTTGTTGGAAGCCGAAG-3'

[0057] The specific upstream and downstream primers of the rhesus monkey GAPDH gene as an internal reference gene are:

[0058] GAPDH F: 5'-AGCCCCATCACCATCTTCC-3'

[0059] GAPDH R: 5'-AATGAGCCCCAGCCTTCTC-3';

[0060] Macaque ABCG2 The primer sequences and fragment sizes for quantification of gene expression levels are...

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Abstract

The present invention provides a kind of RT-qPCR detection rhesus monkey ABCG2 The method of gene transcription level, using the cDNA synthesized by reverse transcription of total RNA extracted from fresh macaque kidney tissue as a template, and using PCR primer combination for real-time fluorescent quantitative PCR amplification, obtained ABCG2 Gene fragments and reference genes GAPDH The Ct value, dissolution peak, and amplification efficiency of the fragments were normalized by routine processing. ABCG2 The ΔΔC(t) value of the gene fold expression, thus obtaining ABCG2 Relative expression values ​​of gene transcript levels. It provides an effective way to study the function of ABCG2 in macaques and the influence of related drugs on it. The invention is suitable for real-time quantitative PCR detection, and has simple operation, high repeatability, low detection cost, high sensitivity and strong specificity.

Description

technical field [0001] The invention relates to a detection method, in particular to a real-time fluorescent quantitative RT-qPCR method for detection of macaque adenosine triphosphate binding cassette transporter ABCG2 The method for gene transcription level belongs to the field of molecular biotechnology. Background technique [0002] In recent years, genome-wide association studies (GWAS) have detected multiple susceptibility loci and related candidate genes that lead to hyperuricemia and gout. in SLC2A9 , SLC22A11 and SLC22A12 Loss-of-function mutations in genes cause hereditary hypouricemia, while overexpression enhances uric acid reabsorption. ABCG2, SLC17A1 and SLC17A3 Defective gene function variants reduce renal and intestinal excretion of uric acid. As a transport protein, ABCG2 is located in the cell membrane and is widely distributed in tissues with secretion and excretion functions. In the kidney, ABCG2 is located in the brush border of the renal pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6851C12Q1/6888
CPCC12Q1/6851C12Q1/6888C12Q2600/166C12Q2600/158C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 唐东红王陈芸叶尤松李哲丽马开利鲁帅尧杨浩陈倩肖涵邱炳玲杨忠
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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