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Standard plasmid molecule for detection of genetic improved soybean strain GTS40-3-2 and constructing method thereof

A technology of genetic improvement and standard plasmids, which is applied in the field of plasmid molecules in the field of bioengineering technology, can solve problems such as the inability to meet the requirements of the genetically modified product labeling system, and achieve accurate results

Inactive Publication Date: 2007-10-31
SHANGHAI JIAO TONG UNIV
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Problems solved by technology

However, its shortcoming is that the plasmid molecule is constructed for the exogenous target gene fragment and can only be used for gene-specific detection of genetically improved crops, which is out of touch with the development trend of detection methods, so it cannot meet the requirements of the genetically modified product labeling system. Require

Method used

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  • Standard plasmid molecule for detection of genetic improved soybean strain GTS40-3-2 and constructing method thereof

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Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1: the construction of standard molecule

[0027] 1. Experimental reagents

[0028] Restriction enzymes Nco I, Xba I, and restriction enzyme buffers were purchased from Shanghai Haojia Technology Development Co., Ltd.;

[0029] pMD18-T vector, T 4 DNA ligase and its buffer were purchased from Shanghai Haojia Technology Development Co., Ltd.;

[0030] dNTPs, Taq DNA polymerase and its buffer, and DL2000 Marker were purchased from Dalian Bao Biological Engineering Co., Ltd.;

[0031] TaqMan probes and primers were synthesized by Shanghai Boya Bioengineering Co., Ltd.

[0032] Other biochemical reagents are imported subpackages or domestic analytical pure.

[0033] 2. Experimental equipment

[0034]DY-501 Nucleic Acid Electrophoresis Apparatus (Shanghai Precision Scientific Instrument Co., Ltd.)

[0035] PTC-100 PCR Amplifier (MJ Research Inc.)

[0036] DNA electrophoresis analysis system, including dark box, digital camera, computer, scanner, inkjet pri...

Embodiment 2

[0053] Example 2: Application of constructed plasmid standard molecules in actual detection

[0054] 1. Experimental materials

[0055] Genetically improved soybean line GTS40-3-2.

[0056] Regular soybean varieties.

[0057] Other genetically improved crop lines: rapeseed GT73, cotton MON531, cotton MON1445, corn GA21, Huafan No. 1 tomato, and corn NK603 lines.

[0058] 2. Enzymes and Reagents

[0059] The extraction and purification of plant genomic DNA used the food DNA extraction kit jointly developed by Shanghai Academy of Agricultural Sciences and Shanghai Entry-Exit Inspection and Quarantine Bureau.

[0060] Plasmid genomic DNA was extracted and purified using a plasmid miniprep kit developed by Jerry Bioengineering (Shanghai) Co., Ltd.

[0061] pMD18-T vector, T 4 DNA ligase and its buffer were purchased from Shanghai Haojia Technology Development Co., Ltd.;

[0062] dNTPs, Taq DNA polymerase and its buffer, and DL2000 Marker were purchased from Dalian Bao Biolo...

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Abstract

The invention discloses a standard plasmid molecule to check genetic improved soybean strain GTS40-3-2 and constructing method, which comprises the following steps: incorporating strain special fragment of the genetic improved soybean strain GTS40-3-2 and special fragment of soybean internal standard gene Lectin; analyzing exogenesis inserting carrier by-pass ortho gene sequence of genetic improved soybean strain GTS40-3-2; designing strain special PCR primer; augmenting; getting strain special fragment of genetic improved soybean strain GTS40-3-2 and soybean internal standard gene special sequence; constructing into a plasmid molecule with molecular cloning method; getting artificial retooling plasmid molecule pMD-RRS. This standard plasmid molecule can be fit for strain special quantitative PCR analysis and check of genetic improved soybean strain GTS40-3-2 sample.

Description

technical field [0001] The invention relates to a plasmid molecule in the technical field of bioengineering, in particular to a standard plasmid molecule for genetically improved soybean strain GTS40-3-2 detection and a construction method thereof. Background technique [0002] The emergence and application of genetic engineering technology has opened up a new era of development for agriculture, medicine and other fields. In 1994, the genetically improved tomato line "FLAVR SAVR" developed by Calgene Corporation using genetic engineering technology was first approved for commercial production in the United States. By 2006, the global planting area of ​​genetically improved crops had reached more than 100 million hectares. At the same time, a large number of genetically improved agricultural products have poured into the Chinese market. In 2004 and 2005, my country's Agricultural GMO Safety Management Office issued safety production licenses for 18 foreign genetically impro...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/29
Inventor 张大兵杨立桃张海波李想郭金超
Owner SHANGHAI JIAO TONG UNIV
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