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New gene MDL1, and quantitative detection method thereof

A technology of MDL1, detection method, applied in genetic engineering, plant genetic improvement, chemical instruments and methods, etc., can solve the problem of molecular mechanism in the primary stage and so on

Inactive Publication Date: 2017-04-19
NANKAI UNIV
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Problems solved by technology

However, understanding of the molecular mechanisms involved in the regulation of mitochondrial replication and expression is still in its infancy

Method used

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  • New gene MDL1, and quantitative detection method thereof
  • New gene MDL1, and quantitative detection method thereof
  • New gene MDL1, and quantitative detection method thereof

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Experimental program
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Embodiment

[0083] (1) Cell culture: lung cancer (NCI-H460), breast cancer (MCF-7), liver cancer (HepG2), cervical cancer (Hela), lymphoma (HTL-90), prostate cancer (PC3) and neuroblastoma The (SH-SY5Y) cell line was cultured in DMEM medium containing 10% fetal bovine serum, the medium was replaced every two days, and the culture medium was subcultured every four days.

[0084] (2) Extraction of total RNA: extract with reference to the kit RNAiso Plus (TaKaRa, Japan), and obtain total RNA with a volume of 25 μL and a concentration of 1.3 μg / μL.

[0085] (3) RNA inversion: 0.8-1 μg RNA was taken out from the total RNA, and RNA inversion was carried out with reference to the kit PrimeScript RT ReagentKit with gDNA Eraser (TaKaRa, Japan). The inversion used a 40-nt long special primer (CTGATCTAGAGGTACCGGATCC-T18 ).

[0086] (4) cDNA amplification: cDNA amplification was carried out with reference to the kit SYBR Premix Ex Taq II (TaKaRa, Japan), and the qRT-PCR instrument used was Mastercyc...

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Abstract

The invention discloses a human mitochondrial new gene MDL1, the full-length nucleotide sequence of a gene MDL 1 AS, and the quantitative detection method of the expression quantity thereof, and belongs to the field of biotechnology. The nucleotide sequence of the gene MDL1 is shown as: 1) a nucleotide sequence shown in SEQ ID NO. 1; or 2) a nucleotide sequence obtained through replacing, deleting or inserting one or several nucleotides in the nucleotide sequence shown in SEQ ID NO. 1. The nucleotide sequence of the gene MDL 1 AS is shown as: 1) a nucleotide sequence shown in SEQ ID NO. 2; or 2) a nucleotide sequence obtained through replacing, deleting or inserting one or several nucleotides in the nucleotide sequence shown in SEQ ID NO. 2. The gene MDL1 and the gene MDL 1 AS are key genes for regulating the gene expression of mitochondrial genes. The mitochondrial energy supply is the basis for various life activities. The present invention verifies that, the expression of the gene MDL1 significantly changes in various tumor cells, so that the gene MDL1 has very important clinical detection and therapeutic value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the full-length sequences of two novel human mitochondrial genes MDL1 and MDL1AS, a detection method and application of the expression levels thereof. Background technique [0002] Animal mitochondria are extremely conserved in evolution, basically including 2 rRNA, 13 mRNA and 22 tRNA coding genes. Mitochondria are the main place for cells to carry out aerobic respiration and provide energy for life activities. The occurrence of various diseases such as tumors requires a large amount of energy supply in a short period of time, so it is inseparable from the abnormal replication and expression of mitochondria. However, understanding of the molecular mechanisms involved in the regulation of mitochondrial replication and expression is still in its infancy. Until 2016, Gao Shan and Bu Wenjun of Nankai University discovered two potential mitochondrial gene expression regulato...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12Q1/68
CPCC07K14/7056C12Q1/6886C12Q2600/158
Inventor 高山姚雪卜文俊阮吉寿程智
Owner NANKAI UNIV
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