Quantitative PCR primer for soybean E2/GIGANTEA homologous gene and application of quantitative PCR primer

A homologous gene and soybean technology, applied in the field of quantitative PCR primers for soybean E2/GIGANTEA homologous genes, can solve the problem of high specificity and specificity of target gene primers

Pending Publication Date: 2020-11-03
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it requires high specificity and specificity in the design of target gene primers, which becomes a limiting factor for accurate analysis of target gene expression levels

Method used

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  • Quantitative PCR primer for soybean E2/GIGANTEA homologous gene and application of quantitative PCR primer
  • Quantitative PCR primer for soybean E2/GIGANTEA homologous gene and application of quantitative PCR primer
  • Quantitative PCR primer for soybean E2/GIGANTEA homologous gene and application of quantitative PCR primer

Examples

Experimental program
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Effect test

Embodiment 1

[0070] Example 1. Design of specific quantitative PCR primers for detecting the expression level of soybean E2 / GIGANTEA homologous genes

[0071] 1. The circadian clock is often composed of multiple genes / proteins, forming multiple interlocked transcription-translation feedback loops. Among them, GIGANTEA (GI) is an important core component. The gene mainly has three homologous genes Glyma.10G221500, Glyma.16G163200 and Glyma.20G170000 in soybean. Among them, Glyma.10G221500 is soybean growth period gene E2, which plays an important role in soybean growth, development and ecological adaptability. Therefore, by adopting the method of real-time quantitative PCR, clarifying the expression characteristics of GIGANTEA (GI) and its homologous genes is very important for understanding and mastering the growth habits of soybean. 10G221500, Glyma.16G163200 and Glyma.20G170000 designed a large number of primers, and obtained a set of PCR primers with good specificity and quantitative ...

Embodiment 2

[0087] Embodiment 2, quantitative PCR primer application

[0088] This example mainly uses the specific quantitative PCR primers prepared in Example 1 to analyze the circadian rhythm characteristics of the expression of soybean E2 / GIGANTEA homologous genes.

[0089] The soybean variety used in this embodiment is Zhonghuang 24 (breeding unit: Institute of Crop Science, Chinese Academy of Agricultural Sciences, the soybean variety that is formed by the selection of Jilin 21 × (Fendou 31 × Zhongdou 19). August 7, 2008 It was approved by the second meeting of the second National Crop Variety Approval Committee on Japan, and the approval number is National Examination Bean 2008003.).

[0090] 1. Plant Zhonghuang 24 until the single leaf is fully unfolded. From zero point, pick the single leaf as a sample every three hours, freeze it through with liquid nitrogen and store it in a -80 degree refrigerator for later use.

[0091] 2. Use the TRNzol kit from Tiangen Company, according t...

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Abstract

The invention discloses a quantitative PCR (Polymerase Chain Reaction) primer for a soybean E2 / GIGANTEA homologous gene and an application of the quantitative PCR primer. The invention provides a setof specific quantitative PCR primers for detecting the expression quantity of the soybean E2 / GIGANTEA homologous gene. The specific quantitative PCR primers are six single-stranded DNA shown as sequences 1-6 in a sequence table. By adopting a real-time quantitative PCR method, the expression characteristics of GIGANTEA (GI) and homologous genes thereof are defined, and the gene not only plays an important role in the soybean growth period, but also plays an important role in circadian movement / circadian clock, so that the gene expression is detected to play an important role in soybean biological research and application. The primer disclosed by the invention has great significance for understanding and mastering the growth habits of soybeans.

Description

technical field [0001] The invention relates to the field of soybean molecular genetic breeding, in particular to a soybean E2 / GIGANTEA homologous gene quantitative PCR primer and application thereof. Background technique [0002] Real-time quantitative PCR technology can conveniently and quickly evaluate the expression level of target genes, and is an important analytical tool widely used in the fields of molecular biology and molecular genetic breeding. However, it requires high specificity and specificity in the design of target gene primers, which becomes a limiting factor for accurate analysis of target gene expression levels. Especially in the case of a large number of copies of homologous genes in the genome, how to design specific primers that can correctly and effectively distinguish each homologous gene is very important. [0003] Soybean is a typical short-day crop and a model material for the study of plant photoperiod response. It is also the most important oil...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/11
CPCC12Q1/6895C12Q1/6851C12Q2600/158C12Q2600/13C12Q2531/113C12Q2545/101
Inventor 蒋炳军孙石韩天富武婷婷袁珊
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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