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Transgenic papaya and detection of transgenic component in transgenic papaya product

A technology for processing products, papaya, applied in the field of product testing, can solve the problem that the internal standard gene of papaya has not been determined, and achieve the effect of eliminating test differences

Inactive Publication Date: 2007-10-03
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the internal standard genes of soybean, corn and rice have been identified, but the internal standard genes of papaya have not been identified.
At present, there is no detection method and detection kit for detecting the content of genetically modified ingredients in transgenic papaya and its processed products

Method used

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  • Transgenic papaya and detection of transgenic component in transgenic papaya product
  • Transgenic papaya and detection of transgenic component in transgenic papaya product
  • Transgenic papaya and detection of transgenic component in transgenic papaya product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0134] Embodiment 1, the preparation of transgenic PRSVR gene antiviral papaya standard reference sample

[0135] In the present invention, DNA is extracted from pure transgenic PRSVR gene antiviral papaya and non-transgenic papaya respectively by CTAB method, and both are diluted to 50ng / ul, and the two kinds of DNA are mixed according to the volume ratio, and the DNA content of transgenic antiviral papaya is prepared. 5%, 2%, 1%, 0.5%, 0.1%, and 0.05% DNA solutions were used as reference samples for quantitative detection.

Embodiment 2

[0136] Embodiment 2, the test kit that detects transgenic PRSVR gene antiviral papaya and its processed products

[0137] The kit contains:

[0138] (1) A pair of specific primers for amplifying the CAR gene:

[0139] Upstream primer 5'AGT GGC TCA ATA TGG TAT TCA CTA CAG A 3'

[0140] Downstream primer 5'AAA ATG TAG ATA TAC CTC CCT TGA GCG 3'.

[0141] 100μl each for 100uM;

[0142] (2) Probes that specifically bind to CAR gene amplification products:

[0143] 5’ATA CTT ACC CAT ATG AGG GAG TGC AAC GTT ATT G 3’

[0144] (3) its specific primer pair for amplifying the PRSVR gene:

[0145] Upstream primer 5'TTG TCC CCT CTT CCG GAG TT 3'

[0146] Downstream primer 5'CTT CCT TGC TTA GAA CGC TTT TCA 3',

[0147] 100μl each for 100uM;

[0148] (4) Probes that specifically bind to PRSVR gene amplification products:

[0149] 5’CCT GGA GTG TAA TGA GGA AGC CAA GAC TTT CTT T 3’.

[0150] (5) its specific primer pair for amplifying the PRSVCP gene:

[0151] Upstream primer 5'TCT...

Embodiment 3

[0161] Embodiment 3, carry out qualitative detection with the kit of embodiment 2

[0162] Proceed as follows:

[0163] (1) Extract the DNA in the cottonseed to be tested with the magnetic bead method DNA extraction kit produced by Promega Company as a template;

[0164] (2) The kit in Example 2 is used for detection. The template DNA is added to the respective reaction systems for amplifying the CAR gene, PRSVCP gene and PRSVR gene, and incubated on a PCR amplification instrument to amplify the CAR gene, PRSVCP gene and PRSVR gene respectively. ;

[0165] Qualitative PCR reaction system (25 μl reaction system) for amplifying CAR gene, PRSVCP gene and PRSVR gene, in which the components are: 10×PCR reaction buffer 2.5 μl, 10×25mM MgCl 2 2.5 μl, 10 mM dNTP 2.5 μl, Taq DNA polymerase 0.2 μl, 5 μM upstream primer 0.5 μl, 5 μM downstream primer 0.5 μl, sterilized ultrapure water 15.3 μl, added template DNA 1 μl (about 30 ng DNA);

[0166] Qualitative PCR cycle parameters are: ...

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Abstract

The present invention discloses one kind of kit for detecting transgenic papaya and transgenic papaya product and its qualitative and quantitative detection method. The quantitative detection method includes the following steps: extracting DNA of the detected sample as the template, and performing PCR amplification with specific primer pair of CAR gene and specific primer pair of exogenous gene; hybridizing the amplified products separately with the probe and detecting the hybridization generated detectable signal; and analyzing the detected data to detect the transgenic component content in papaya and papaya product quantitatively. The said method of the present invention is accurate and simple.

Description

technical field [0001] The invention relates to product detection technology, more specifically, the invention relates to a detection kit and a detection method for qualitatively and quantitatively detecting transgenic papaya and its processed products. Background technique [0002] In recent years, with the development of biotechnology, genetically modified crops have been rapidly commercialized and entered the international market through trade. The development of biotechnology has significantly improved the yield and quality of agricultural products, and alleviated the impact of population growth, scarcity of natural resources, reduction of arable land area and unpredictable natural disasters to a certain extent. However, the safety of genetically modified crops has aroused great concern. [0003] Many countries in the world, such as the European Union, Japan, Australia, Singapore and other countries, have adopted legislation or other forms to strictly manage genetically...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 许文涛黄昆仑罗云波郭峰张方方
Owner CHINA AGRI UNIV
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