Accurate and efficient eukaryotic biological gene identification method based on transcriptome

A technology of eukaryotic organisms and identification methods, which is applied in the field of eukaryotic gene identification, can solve the problems of increasing economic costs and time costs, and achieve the effect of complete and compact technical design, reasonable design, and meeting research needs

Inactive Publication Date: 2018-11-06
SICHUAN AGRI UNIV
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Problems solved by technology

Greatly increased the economic cost and time cost of identification

Method used

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Embodiment

[0023] An accurate and efficient eukaryotic gene identification method based on transcriptome, comprising the following process steps:

[0024] The first step is to identify the prediction of gene expression: use THE HUMAN PROTEIN ATLAS database to search for the tissue expression of Leptin, and find that its expression level is high in soft tissue and adipose tissue. Only the Dabry's sturgeon over 50cm can easily collect adipose tissue, and the Dabry's sturgeon in the laboratory sample is about 20cm. Considering that the liver contains a large number of fat cells, it is speculated that leptin is also expressed in the liver, so the Dabry's liver was collected. As a transcriptome sequencing sample;

[0025] Step 2: Collection of high-expression samples of genes to be identified, RNA extraction and reverse transcription: quickly dissect Dabry's sturgeon in a second-level biological safety cabinet to take out the liver, freeze it in liquid nitrogen, extract RNA, and conduct agaro...

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Abstract

The invention discloses an accurate and efficient eukaryotic biological gene identification method based on a transcriptome. The method comprises 1, quasi-identification gene expression prediction, 2,acquisition of a quasi-identification gene high expressed sample, RNA extraction and reverse transcription, 3, transcriptome sequencing, 4, transcriptome data analysis and storage, 5, transcriptome data retrieval and 6, gene identification primer design and gene identification. Through the close specie research report analysis, the transcriptome of the high expressed sample of the gene to be identified is determined, the target sequence of the gene to be identified is screened in the transcriptome, and the gene is identified and cloned based on the target sequence so that an accurate gene sequence is obtained. The method is suitable for identification of functional genes of most eukaryotes, is very accurate, satisfies the research demands of the functional gene identification in the research process, has a reasonable design, is easy to use, realizes a low cost and is suitable for promotion and use.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an accurate and efficient eukaryotic gene identification method based on transcriptome. Background technique [0002] Since the completion of the human genome, the genomes of more and more organisms have been sequenced. Genome-based identification of biological genes is relatively easy. However, the proportion of species whose genomes have been sequenced is very small, and most species need to identify their own genes through homologous cloning. Homologous cloning works better when the corresponding genes have been identified in closely related species. However, homologous cloning often fails to achieve satisfactory results in the identification of genes with low consistency among species and genes of some species with special evolutionary status. It is often necessary to analyze the conserved sites of the gene and design an incorporation primer, which may require several or even ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6869G06F19/18
CPCC12Q1/6869C12Q1/6888C12Q2535/122
Inventor 李志琼汪斌陈虎齐锦雯陈德芳唐妮王书瑶田正志吴源冰王美徐少奇
Owner SICHUAN AGRI UNIV
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