Tobacco Heat Shock Protein Gene and Its Application

A technology of heat shock protein and tobacco, which is applied in application, genetic engineering, plant gene improvement, etc., can solve the problems of not constant expression and different stability, and achieve the effect of improving accuracy

Active Publication Date: 2017-08-22
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, previous research results have shown that the stability of housekeeping genes in samples of different tissues and organs and developmental stages is different, and their expression is not constant

Method used

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  • Tobacco Heat Shock Protein Gene and Its Application
  • Tobacco Heat Shock Protein Gene and Its Application
  • Tobacco Heat Shock Protein Gene and Its Application

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0022] A tobacco heat shock protein gene, the base sequence of which is shown in SEQ ID NO:1.

[0023] The amino acid sequence encoded by the tobacco heat shock protein gene is shown in SEQ ID NO:2.

[0024] The application of quantitative expression analysis of tobacco heat shock protein gene in tobacco real-time fluorescent quantitative PCR, PCR primers are designed for the tobacco heat shock protein gene, and quantitative expression analysis based on real-time fluorescent quantitative PCR experiment is carried out in tobacco. .

[0025] The application of the quantitative expression analysis of tobacco heat shock protein gene in tobacco real-time fluorescent quantitative PCR, the specific primer pair of tobacco real-time fluorescent quantitative PCR is SEQ ID NO: 3 and SEQ ID NO: 4.

[0026] The commonly used internal reference genes in tobacco are 8 housekeeping genes, including Actin, PP2A, L25, Ntubc2, EF-1α, α-Tubulin, β-Tubulin and 18S rRNA. Based on chip data and re...

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Abstract

The invention discloses a tobacco heat shock protein gene and the base sequence of the tobacco heat shock protein gene is shown in SEQIDNO: 1. According to the invention, based on real-time fluorescent quantitative PCR experiment carried out on 44 tissue samples of tobacco at all development stages, the expression stability of HSC70-1 gene and the expression stability of the frequently used 8 reference genes of the tobacco are compared to obtain a comparison result that the expression stability of HSC70-1 gene is superior to that of all the 8 frequently used reference genes. Therefore, when HSC70-1 gene is applied to the real-time fluorescent quantitative PCR experiment of the tobacco, the accuracy of the relatively quantitative result can be improved and a new tool is provided for the real-time fluorescent quantitative PCR experiment of the tobacco.

Description

technical field [0001] The invention relates to the selection of internal reference genes in tobacco real-time fluorescent quantitative PCR experiments, in particular to the application of tobacco heat shock protein gene HSC70-1 as a new internal reference gene in real-time fluorescent quantitative PCR. Background technique [0002] Real-time fluorescence quantitative PCR is an important research method for gene expression research, which has the characteristics of accurate quantification, high sensitivity, and good repeatability. In order to eliminate the impact of factors such as template quality / quantity, PCR reaction efficiency, and experimental material differences on quantitative results, real-time fluorescent quantitative PCR usually uses internal reference genes for calibration and standardization, and the accuracy of the results largely depends on the selection of internal reference genes. . [0003] Housekeeping genes are usually used as internal reference genes i...

Claims

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Application Information

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Patent Type & AuthorityPatents(China)
IPC IPC(8): C07K14/415C12N15/29C12Q1/68
CPCC07K14/415C12Q1/6895C12Q2600/158C12Q2600/166
Inventor曹培健王燃许亚龙李泽锋卢鹏杨军李锋魏春阳罗朝鹏金立锋林福呈
OwnerZHENGZHOU TOBACCO RES INST OF CNTC