Endogenous reference gene suitable for brachypodium distachyon exogenous gene detection and copy number analysis, and application thereof

A kind of technology of Brachypodium erinaceus and internal standard gene, which is applied in the field of bioengineering

Inactive Publication Date: 2014-01-22
SHANGHAI CHENSHAN BOTANICAL GARDEN
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no report on the internal standard gene suitable for the detection of exogenous genes and copy number analysis of Brachypodium distachyon in the world.

Method used

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  • Endogenous reference gene suitable for brachypodium distachyon exogenous gene detection and copy number analysis, and application thereof
  • Endogenous reference gene suitable for brachypodium distachyon exogenous gene detection and copy number analysis, and application thereof
  • Endogenous reference gene suitable for brachypodium distachyon exogenous gene detection and copy number analysis, and application thereof

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Experimental program
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Effect test

Embodiment 1

[0031] 1. Experimental materials

[0032] 1.1 Plant material

[0033] Brachypodium distachyon: The selected materials of Brachypodium distachyon are diploid plants and polyploid plants, see Table 1 for details.

[0034] Brachypodium distachydenta material selected in the experiment of table 1

[0035]

[0036]

[0037] Other common crops: 1. Rose; 2. Cotton; 3. Ryegrass; 4. Carnation; 5. Ophiopogon japonicus; 6. Gerbera; 7. Bermudagrass; 8. Tobacco; 9. Corn; 10. Rice; 11. Soybean; 12. Wheat; 13. Barley; 14. Arabidopsis.

[0038] 1.2 Enzymes and Reagents

[0039] Restriction enzymes, dNTPs, DL2000Markers, Taq DNA polymerase and their buffers were purchased from Takara. Primers were synthesized by a bioengineering company, and other biochemical reagents were imported or domestically produced.

[0040] 1.3 Experimental Instruments

[0041] EPS-300 nucleic acid electrophoresis instrument (Shanghai Tianneng Technology Co., Ltd.), T100 PCR amplification instrument (Bio-R...

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Abstract

The invention discloses an endogenous reference gene suitable for brachypodium distachyon exogenous gene detection and copy number analysis, and application thereof. Through bioinformatics analysis and molecular biology experimental verification, a BDFIM gene serving as the endogenous reference gene for transgenic brachypodium distachyon qualitative and quantitative polymerase chain reaction (PCR) detection and copy number analysis is found from the brachypodium distachyon, and the nucleotide sequence of the gene is as shown in SEQ ID NO.1. The endogenous reference gene can be applied to qualitative and quantitative detection of the exogenous gene of the transgenic brachypodium distachyon, and also can be applied to analysis on the copy number of the exogenous gene in the transgenic brachypodium distachyon species.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to an internal standard gene suitable for detection and copy number analysis of exogenous genes of transgenic Brachypodium distachyon and its application. Background technique [0002] The so-called internal standard gene is a type of gene in a certain plant that has the characteristics of inter-species specificity, intra-specific non-specificity, and low copy number. Interspecies specificity means that the selected gene is specific to this species, but has very low homology in other species; intraspecific nonspecificity means that the internal standard gene has different homology in different cultivars of the same species. High homology and similarity. For example, in the quantitative detection of plant genetically modified products, the internal standard gene is often used as the internal control of quantitative detection, and the internal standard gene is required to have t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 储昭庆朱宏
Owner SHANGHAI CHENSHAN BOTANICAL GARDEN
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