RT-qPCR detection method of slc2a9/glut9 gene transcription level in macaques

A technology of gene transcription and rhesus monkey, which is applied in biochemical equipment and methods, microbiological determination/inspection, etc., to achieve the effect of simple method, high sensitivity and high repeatability

Active Publication Date: 2022-02-11
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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  • Abstract
  • Description
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Problems solved by technology

At present, there is no use of rhesus monkeys at home and abroad. SLC2A9 / Study on Quantitative Detection Method of GLUT9 Transcript Level

Method used

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  • RT-qPCR detection method of slc2a9/glut9 gene transcription level in macaques
  • RT-qPCR detection method of slc2a9/glut9 gene transcription level in macaques
  • RT-qPCR detection method of slc2a9/glut9 gene transcription level in macaques

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Embodiment

[0052] A RT-qPCR detection in macaques SLC2A9 / The method of GLUT9 gene transcription level, comprises the following steps:

[0053] (1) Design the following primers:

[0054] According to the macaque in the NCBI gene bank ( Macaca Mulatta ) monkey SLC2A9 / GLUT9 nucleotide sequence number NM--020041.2, macaque ( Macaca Mulatta )of GAPDH Nucleotide sequence number NM--001195426.1 GAPDH As an internal reference gene design, primers were designed with rhesus monkeys, gene accession number NM_001195426.1, synthesized by Shanghai Bioengineering Co., Ltd.:

[0055] Macaque SLC2A9 The specific upstream and downstream primers for the expression level of / GLUT9 gene are:

[0056] SLC2A9 / GLUT9 F:5'-CCACGCTACCTGCTCTTGGA-3'

[0057] SLC2A9 / GLUT9 R: 5’-TGCTTTACCCAAGAACGTTTGGA-3’

[0058] The specific upstream and downstream primers of the rhesus monkey GAPDH gene as an internal reference gene are:

[0059] GAPDH F: 5'-AGCCCCATCACCATCTTCC-3'

[0060] GAPDH R: 5'-AA...

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Abstract

The present invention provides a kind of RT-qPCR detection rhesus monkey SLC2A9 / GLUT9 gene transcription level method, the total RNA extracted from macaque fresh kidney tissue was reverse-transcribed into cDNA as a template, and real-time fluorescent quantitative PCR amplification was performed using PCR primer combination to obtain SLC2A9 / GLUT9 gene fragment and reference gene GAPDH The Ct value and dissolution peak of the fragment; the homogenized SLC2A9 / ΔΔC(t) value of multiple expression of GLUT9 gene, obtained SLC2A9 The relative expression value of / GLUT9 gene transcription level provides an effective way for studying the function of macaque glucose transporter 9 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 present invention relates to a detection method, in particular to a real-time fluorescent quantitative qPCR method for detection of rhesus monkey glucose transporter 9 SLC2A9 The invention relates to a method for transcription level of GLUT9 gene, which 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. GLUT9 is a fructose transporter that belongs to SLC2A9 The family, located on the basolateral membrane of renal tubular epithelial cells, is mainly responsible for the secretion of uric...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2561/113C12Q2563/107C12Q2545/114C12Q2521/107C12Q2545/101
Inventor 唐东红王陈芸叶尤松李哲丽马开利鲁帅尧杨浩肖涵邱炳玲陈倩杨忠
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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