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Isothermal amplification assay kit for phosphomannose isomerase

A technology of mannose phosphate and isothermal amplification, which is applied in the detection of mannose phosphate isomerase, and the warm amplification method is used to detect mannose phosphate isomerase gene, which can solve the problem of false negative results of detecting target fragments and DNA fragment fragmentation and other problems, to achieve the effect of increasing the positive detection rate, reducing false negative results, and having a wide range of applications

Inactive Publication Date: 2011-06-15
ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
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AI Technical Summary

Problems solved by technology

Because most of the food is deeply processed, the DNA fragments are seriously broken, and the detection target fragments are too long to produce false negative results.

Method used

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  • Isothermal amplification assay kit for phosphomannose isomerase
  • Isothermal amplification assay kit for phosphomannose isomerase

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Experimental program
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Effect test

Embodiment 1

[0066] Samples: Known transgenic corn events 3272, MIR604, MON89034, BT176, 1507, MON810, MON863, NK603, GA21, BT21, of which 3272 and MIR604 were positive for phosphomannose isomerase, and the primer pairs used for detection were:

[0067] phosphomannose isomerase

[0068] Primer 1: 5-TCCCGATTCCAGTGGATGA

[0069] Primer 2: 5-ACAGTCACCGGTGATTCGT

[0070] Primer 3: 5-ACAAAATGGCGGCACTCTGCT-GCCTTCTCGCTGCATGAC

[0071] Primer 4: 5-CGTCGAAGGCGATGCAACGT-GGCGGCAATAAACGCTGAT

[0072] The reaction system is as follows:

[0073] Table 1 Phosphomannose Isomerase Detection System 1

[0074] Working fluid name

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Abstract

The invention relates to an isothermal amplification assay kit for phosphomannose isomerase, belonging to the field of food inspection. The invention discloses a primer for amplifying the phosphomannose isomerase, reagent for amplifying the phosphomannose isomerase and a preparation method for an amplification product. The isothermal amplication assay kit has the advantages of high sensitivity and high accuracy, is suitable to screen transgene ingredients in corn, beet, wheat, rice, cassava, sweet oranges and grapes and is convenient to popularize and apply in various detection mechanisms.

Description

technical field [0001] The invention provides a method for detecting mannose phosphate isomerase, which belongs to the field of genetically modified food detection, and in particular relates to a technique for detecting mannose phosphate isomerase gene by using a loop-mediated isothermal amplification method. Background technique [0002] The transformation frequency of plant genetic transformation is usually low, and selectable markers must be used in selection to ensure the survival of transformed cells and prevent the regeneration of non-transformed cells. Traditional screening methods use antibiotic and herbicide-like selective agents, but the impact of these substances on humans and the environment is not fully understood, so non-antibiotic screening systems have emerged. [0003] The mannose positive selection system belongs to one of the non-antibiotic screening systems. It uses the phosphomannose isomerase gene (pmi) of Escherichia coli as a selection gene and mannos...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 贺艳郑文杰朱水芳刘伟刘跃庭张裕君廖芳
Owner ANIMAL & PLANT & FOOD INSPECTION CENT OF TIANJIN ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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