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Monitoring of Degaradation of CP4-EPSPS Protein, Endogenous and Exogenous Gene in Roundup Ready Soybean during Food Processing

Author: WuHongHong
Tutor: ShenWenZuo
School: Nanjing Agricultural College
Course: Biochemistry and Molecular Biology
Keywords: Roundup Ready soybean Processing CP4-EPSPS protein and its gene degradation Exogenous mimic experiment
CLC: S565.1
Type: Master's thesis
Year: 2011
Downloads: 8
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Abstract

Roundup Ready soybean is a line that transforming Cp4-epsps gene into a wild type soybean, which has higher resistance to the glyphosate than the wild type. Compared with the wild type soybean, Roundup Ready soybean is easy to manage and can reduce the production cost. Accordingly, it is farmed quickly and widely.As we all known, people’s concentration on the safty of GMOs is growing.As the main transgenic crops, researcher’s interest on the Roundup Ready soybean is rising as well. However, current studies are mainly on the detection methodology; the detection during food processing is lacking.The research in this paper shed light on the physics, chemicals and biological factors involved in protein and gene degradation in Roundup Ready soybean during processing. Applying PCR and Western blot technology; we investigated the degradation of CP4-EPSPS protein and its gene, and then conduct some exogenous mimic experiments to verify it.In this study, we found that degradation of CP4-EPSPS protein and its gene in Roundup Ready soybean during processing are associated with the intensity of treatment. Similarly, exogenous mimic experiments showed that different treatments (heating, microwaving and autoclaving) lead to different results. The results are also dependent with the intensity of treatment.Using Western blot method, the research showed 219-233 aa area (DADGVRTIRLEGRGK) is more stable than the 19-33 aa area (SGTVRIPGDKSISHR) in CP4-EPSPS protein.Furthermore,843-1123 bp fragment and 1144-1305 bp fragment are the most stable fragments in Cp4-epsps gene and lectin gene respectively. Similar result was also exhibited in the exogenous mimic experiments.Hydrolase treatments such as pancreatin, pepsin, trypsin, papain etc. in the exogenous mimic experimentssuggested that hydrolase plays a certain role in the degradation of CP4-EPSPS protein and its gene in Roundup Ready soybean during food processing. The results demonstrated both 219-233 aa polypeptide (DADGVRTIRLEGRGK) and 19-33 aa polypeptide (SGTVRIPGDKSISHR) of CP4-EPSPS protein in transgenic Roundup Ready soybean are digested totoally when treated with the 8000 U/g substrate papain and pepsin (opposite results were detected when treated with trypsin and RNase A). However, the 219-233 aa polypeptide is more stable than 19-33 aa polypeptide when treated with pancreatin. In addition, PCR technique also could detect the degradation of endogenous and exogenous gene of transgenic Roundup Ready soybean. It maybe associated withthe exposure of DNA due to the hydrolase digest the binding protein of DNA which contributes to its stability. Accordingly, it is suggested that nuclease, corporate with certain factors, can cause the degradation of endogenous and exogenous gene of transgenic Roundup Ready soybean.To sum up, the degradation of CP4-EPSPS protein and its gene in Roundup Ready soybean is complicated during processing, which related to physics, chemicals and biologicals factors. The research not only complements the basic reasearch of transgenic food, but also provides somemore convinced results and effective technology to detect transgene components in foodstuff products in market.

Monitoring of Degaradation of CP4-EPSPS Protein, Endogenous and Exogenous Gene in Roundup Ready Soybean during Food Processing

Abstract

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