Gene chip detection method of transgenic maize

Abstract

The invention discloses a gene chip detection method of transgenic maize. The main steps comprise primer and probe determination, chip preparation, PCR (polymerase chain reaction) amplification, PCR product marking, chip hybridization, chip scanning and data processing. By adopting the detection method, sequences of seven main exogenous DNAs (deoxyribonucleic acid) commonly used in transgenic maize, such as promoters, terminators, insect-resistant genes, glyphosate-resistant genes and the like, can be detected and analyzed through one step, so that the detection method of transgenic maize has the advantages of high throughput, miniaturization, automation, rapidity, high efficiency and high sensitivity, and can be widely applied to related fields such as import-export inspection and quarantine, and the like.

Description

technical field [0001] The invention relates to a detection method, specifically a method for detecting transgenic corn products by using a gene chip. Background technique [0002] With the rapid development of genetic engineering technology, genetically modified crops have entered the stage of large-scale commercial production. There are more than 100 varieties of genetically modified crops, mainly including soybeans, corn, cotton, rape, etc., and genetically modified foods and food ingredients processed and produced from genetically modified crops There are more than 4000 species. However, while genetically modified crops bring huge economic benefits, there are also many potential problems. Since there is still no scientific basis to prove the safety of genetically modified products to human health and the environment, the potential risks of genetically modified products to health and the environment have aroused great concern and widespread concern from governments and t...

AI Technical Summary

Problems solved by technology

But it also has its limitations: (1) The detection range is narrow, and the genetically modified food needs to contain the range to be detected, such as EPSPS gene herbicide-tolerant soybeans, Bt gene-transformed insect-resistant corn, etc., so it can only be tested in commercial GMF varieties. use in the case of

At present, commercial ELISA kits can only detect a few kinds of GMF, and a kit is only aimed at a specific transgenic product, so it cannot detect food samples with multiple mixed components with high throughput and fast; (2) it is prone to false positives. Negative results. On the one hand, the content of "new protein" in genetically modified products is usually very low, which is difficult to detect. On the other hand, proteins are volatile during food processing. Proteins in processed foods are likely to lose the antigenic epitopes targeted by antibodies. resulting in false negative ELISA test results

In addition, new modifications of proteins before and after expression in the recipient organism's genome can also lead to reduced detection sensitivity and false negative results

[0013] As far as the ELISA and PCR technologies commonly used in the detection of genetically modified products are concerned, the biggest disadvantage is the narrow detection range, low efficiency, and the inability to simultaneously detect multiple samples with high throughput and large scale, especially in the case of ignorance of the genetically modified background. Under these conditions, it is almost impossible to screen each candidate gene sequence or protein one by one.