Screening and application of fluorescent quantitative reference genes of pear fruits at different development stages

A technology of real-time fluorescence quantification and internal reference gene, which is applied in the field of molecular biology research, can solve the problems that the gene expression level cannot be truly reflected, the expression cannot be stabilized, and the conclusion is opposite, so as to achieve the data authenticity and reliability, high accuracy, and application wide range of effects

Inactive Publication Date: 2017-09-22
NANJING AGRICULTURAL UNIVERSITY
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Problems solved by technology

However, more and more reports point out that under different growth stages or experimental conditions, the results of traditional housekeeping genes used as internal reference ge

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  • Screening and application of fluorescent quantitative reference genes of pear fruits at different development stages
  • Screening and application of fluorescent quantitative reference genes of pear fruits at different development stages
  • Screening and application of fluorescent quantitative reference genes of pear fruits at different development stages

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Embodiment Construction

[0048] The present invention studies the internal reference genes for fruit development of the three main pear varieties Cuiguan, Fengshui and Xueqing. Mainly take Cuiguan (cg) pear as example below, in conjunction with accompanying drawing and embodiment describe in detail:

[0049] 1. Based on the research on genes related to fruit development, four stages of fruits in the whole development process of Cuiguan, Fengshui and Xueqing were selected for transcriptome sequencing. Nine candidate internal reference genes with high expression levels and relatively stable expression levels were screened.

[0050] 2. Select 9 genes with relatively stable expression from the RNA-Seq expression profile database as candidate internal reference genes (Table 2), and the pear housekeeping gene tubulin gene (PbrTUB) as a control. According to the cDS sequence of candidate internal reference genes and the design principles of qRT-PCR, 10 pairs of specific primers were designed using Primer5.0...

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Abstract

The invention discloses screening and application of fluorescent quantitative reference genes of pear fruits at different development stages, and relates to the field of pear molecular biology researches. Reference genes, which are more stable than common housekeeping genes at different development stages of Cuiguan, Fengshui and Xueqing pear fruits, are pertinently obtained according to dynamic RNA (Ribonucleic Acid)-Seq sequencing results, post-period experience validation and data analysis of pear fruits of various varieties; the difference between samples and the difference in the samples, which are caused by the selection of the reference genes, are reduced to the greatest extent; the correction and standardization of a target gene expression level can be realized, and accurate quantification of functional gene expression of the pear fruits is realized.

Description

technical field [0001] The invention relates to the field of molecular biology research, in particular to the screening and application of real-time fluorescent quantitative PCR internal reference genes during pear fruit development. Background technique [0002] Pear (Pyrus) is one of the main fruit trees in my country. It has a long history of cultivation. The cultivation area covers all parts of the country. The cultivation area and output rank first in the world. In recent years, Chinese researchers have carried out many studies on pears from the molecular biology level, especially after the completion of the world's first pear whole genome sequencing, assembly and gene annotation in May 2012, gene expression analysis has been widely used to reveal the pear fruit. In the study of gene regulation mechanism. [0003] Real-time fluorescent quantitative PCR has the characteristics of high sensitivity, good repeatability, strong specificity and high throughput, and is curren...

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6851C12Q1/6895C12Q2600/158C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 谷超王国明张绍铃郝萍萍郭志华金子明
Owner NANJING AGRICULTURAL UNIVERSITY
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