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Primer set and probe for accurate identification of two-generation soybean product transgenic epsps gene and its identification method

A primer set and soybean technology, which is applied in the field of biotechnology analysis, can solve the problem of not being able to identify the second-generation epsps gene in soybean products in real time, and achieve the effects of scientific and reasonable detection method, simple process and high sensitivity

Inactive Publication Date: 2018-02-23
四川省农业科学院分析测试中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The technical problem to be solved by the present invention is to provide a primer set and probe for real-time identification of two generations of soybean products trans epsps gene and a method for detecting whether soybean products contain trans epsps gene, which mainly solves the problem that the existing technology cannot identify soybean products in real time. Problems with the second generation of transgenic epsps

Method used

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  • Primer set and probe for accurate identification of two-generation soybean product transgenic epsps gene and its identification method
  • Primer set and probe for accurate identification of two-generation soybean product transgenic epsps gene and its identification method
  • Primer set and probe for accurate identification of two-generation soybean product transgenic epsps gene and its identification method

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Embodiment

[0028] Such as figure 1 Shown, the present invention mainly detects at the first generation and the second generation trans epsps gene in the soybean product, and concrete method is as follows:

[0029] First, the present invention designs a set of primer sets and probes for amplifying DNA in soybean products on a fluorescent quantitative PCR instrument. The probe sequence is: 5'-FAM-TCATCGAGCCGATCATGACGCG-TAMARA-3';

[0030] The primer set includes an upstream primer and a downstream primer, the sequences of which are as follows:

[0031] Upstream primer: 5'-ACACGCCCGGCATCAC-3';

[0032] Downstream primer: 5'-CTGCAGCATCTTTTCCGTATGA-3'.

[0033] Then, when it is necessary to detect whether the soybean product contains the trans epsps gene, it is mainly divided into two steps, as follows:

[0034] (1) Prepare identification reagents: 25.0 μL PCR reaction system, 2 μl of tested soybean DNA dilution solution with a content of 25 ng / μL, the 25.0 μL PCR reaction system includes...

example 1

[0042] Such as figure 2 As shown, the primer sets and probes designed by the present invention are used for known transgenic corn, transgenic rice, first-generation and second-generation transgenic epsps gene soybeans, transgenic rape samples and non-transgenic corn, rice, rape, soybean samples testing, according to figure 2 From the results shown, it can be seen that the primer set and probe designed in the present invention can only detect the first-generation and second-generation epsps genes in soybean products, and the specificity is extremely high.

example 2

[0044] Such as image 3 and 4 As shown, the present invention is used to test the 5 copies of epsps gene DNA fragments contained in the system, and the test data are as shown in Table 1; 40 times of repetitions using the present invention to contain 5 copies of the epsps gene DNA fragments contained in the reaction system at a 100% confidence level The epsps gene DNA fragment was tested, and the test data are shown in Table 2. according to image 3 As can be seen from the shown results, the epsps gene DNA fragment containing 5 copies in the system can be accurately identified by using the present invention, according to Figure 4 From the results shown, it can be seen that DNA fragments of the epsps gene were detected 40 times in total for 40 repeated tests.

[0045] The sensitivity test result of table 1 the inventive method

[0046]

[0047] Sensitivity (lower detection limit 5copies) test data of the inventive method under the 100% confidence level of table 2

[004...

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Abstract

The invention discloses a primer group and a probe used for identifying trans-epsps genes of soybean products of two generations in real time, and a method for detecting whether the soybean products contain the trans-epsps genes or not, and solves the problem that the second-generation trans-epsps gene of the soybean product cannot be detected in real time in the prior art. The primer group comprises upstream and downstream primers; a sequence of the upstream primer is 5'-ACACGCCCGGCATCAC-3'; a sequence of the downstream primer is 5'-CTGCAGCATCTTTTCCGTATGA-3'; and a sequence of the probe is 5'-FAM-TCATCGAGCCGATCATGACGCG-TAMARA-3'. The method for detecting the trans-epsps genes by utilizing the primer group and the probe comprises the steps of preparing a PCR system and a tested soybean DNA dilute solution; and adding the tested soybean DNA dilute solution into the PCR system for performing an amplified reaction.

Description

technical field [0001] The invention relates to the field of biotechnology analysis methods, in particular to a primer set and a probe for real-time identification of two generations of soybean products trans epsps gene and a method for detecting whether soybean products contain trans epsps gene. Background technique [0002] The first generation of epsps genetically modified soybeans (GTS40-3-2) was successfully developed and commercialized in the 1990s. my country has approved the import of soybeans for raw material processing since 2004; the second generation of epsps genetically modified soybeans (MON89788) was released in 2008. It has been approved to be imported by our country for raw material processing. The epsps gene sequences in the first and second generation transgenic soybean products are not exactly the same, only 81% homology. The previous method or standard can only identify the first generation epsps gene transgenic soybean but not the second generation epsp...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6851C12N15/11
CPCC12Q1/6851C12Q1/6895C12Q2600/158C12Q2531/113C12Q2545/113C12Q2563/107
Inventor 宋秋池
Owner 四川省农业科学院分析测试中心
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