scholarly journals Recombinase Polymerase Amplification (RPA) of CaMV-35S Promoter and nos Terminator for Rapid Detection of Genetically Modified Crops

2014 ◽  
Vol 15 (10) ◽  
pp. 18197-18205 ◽  
Author(s):  
Chao Xu ◽  
Liang Li ◽  
Wujun Jin ◽  
Yusong Wan
Keyword(s):  
Rapid Detection ◽  
Genetically Modified ◽  
Genetically Modified Crops ◽  
Recombinase Polymerase Amplification ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Camv 35S ◽  
Nos Terminator

scholarly journals A simple and accurate PCR method for detection of genetically modified rice

2019 ◽  
Vol 17 (2) ◽  
pp. 847-851 ◽  
Author(s):  
Payam Safaei ◽  
Ebrahim Molaee Aghaee ◽  
Gholamreza Jahed Khaniki ◽  
Setareh Agha Kuchak Afshari ◽  
Sassan Rezaie
Keyword(s):  
Genetically Modified ◽  
Sucrose Phosphate Synthase ◽  
Cauliflower Mosaic Virus ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Rice Varieties ◽  
Camv 35S ◽  
Nos Terminator ◽  
Gm Rice ◽  
Pcr Method

Abstract Background Legislation regulating for labeling and use of genetically modified (GM) crops are increased considerably worldwide in order to health and safety assurance of consumers. For this purpose, a polymerase chain reaction (PCR) method has been developed for detection of GM rice in people’s food diet. Methods In this study, eighty-one non-labeled rice samples were collected randomly from different market sites of Tehran, Iran. In order to analysis, rice genomic DNA was extracted using MBST DNA extraction kit and subsequently, sucrose phosphate synthase (SPS) gene was used to confirm the quality of extracted DNA. Then, cauliflower mosaic virus (CaMV) 35S promoter and Agrobacterium nopaline synthase (NOS) terminator were selected as screening targets for detection of GM rice sequences by PCR. Results According to our results, 2 out of 81 (2.4%) samples tested were positive for CaMV 35S promoter while no positive result was detected for NOS terminator. Conclusion The obtained data indicated that this method is capable to identify the GM rice varieties. Furthermore, it can demonstrate the possibility of the presence of GM rice in Tehran’s market, thus putting emphasis on the requirement for developing a precise approach to evaluate this product.

scholarly journals Semi-quantitative detection of genetically modified grains based on CaMV 35S promoter amplification

2000 ◽  
Vol 3 (2) ◽  
Author(s):  
Alejandro C. Tozzini ◽  
M. Carolina Martínez ◽  
M. Florencia Lucca ◽  
Cecilia Vázquez Rovere ◽  
Ana Julia Distéfano ◽  
...  
Keyword(s):  
Genetically Modified ◽  
Camv 35S Promoter ◽  
Quantitative Detection ◽  
35S Promoter ◽  
Camv 35S

Aberrant promoter methylation occurred from multicopy transgene and SU(VAR)3 - 9 homolog 9 (SUVH9) gene in transgenic Nicotiana benthamiana

2012 ◽  
Vol 39 (9) ◽  
pp. 764 ◽  
Author(s):  
Gi-Ho Lee ◽  
Seong-Han Sohn ◽  
Eun-Young Park ◽  
Young-Doo Park
Keyword(s):  
Gene Silencing ◽  
Nicotiana Benthamiana ◽  
Fluorescent Protein ◽  
Cauliflower Mosaic Virus ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Camv 35S ◽  
Histone Deacetylase 1 ◽  
Transgenic Lines ◽  
Green Fluorescent

The chemical modification of DNA by methylation is a heritable trait and can be subsequently reversed without altering the original DNA sequence. Methylation can reduce or silence gene expression and is a component of a host’s defence response to foreign nucleic acids. In our study, we employed a plant transformation strategy using Nicotiana benthamiana Domin to study the heritable stability of the introduced transgenes. Through the introduction of the cauliflower mosaic virus (CaMV) 35S promoter and the green fluorescent protein (GFP) reporter gene, we demonstrated that this introduced promoter often triggers a homology-dependent gene-silencing (HDGS) response. These spontaneous transgene-silencing phenomena are due to methylation of the CaMV 35S promoter CAAT box during transgenic plant growth. This process is catalysed by SU(VAR)3–9 homologue 9 (SUVH9), histone deacetylase 1 (HDA1) and domains rearranged methylase 2 (DRM2). In particular, we showed from our data that SUVH9 is the key regulator of methylation activity in epigenetically silenced GFP transgenic lines; therefore, our findings demonstrate that an introduced viral promoter and transgene can be subject to a homology-dependent gene-silencing mechanism that can downregulate its expression and negatively influence the heritable stability of the transgene.

scholarly journals Screening of genetically modified Corn Zea mays in Iraqi market

2013 ◽  
Vol 7 (1) ◽  
pp. 21-29
Author(s):  
Ghaith Lotfi Aarif ◽  
Bilal Kamil Sulaiman ◽  
Zahra M. Alkhafaji
Keyword(s):  
Zea Mays ◽  
Multiplex Pcr ◽  
Animal Feed ◽  
Genetically Modified ◽  
Protein A ◽  
Genetically Modified Crops ◽  
Specific Gene ◽  
Maize Crop ◽  
Nos Terminator ◽  
The Government

Detection of the genetically modified crops could be done by screening certain markers usually used in modification. In this study polymerase chain reaction (PCR) technology was used to investigate the presence of the promoter P35s and nos terminator in the genetically modified corn Zea mays. 72 samples of the maize crop collected from inside Iraqi market from various sources, including imported crops and other local strains used for agriculture or for the production of animal feed. DNA extracted from the corn seeds by two methods, the efficiency of extraction was compared between the two procedures, the purity of DNA samples extracted ranged between 1.4- 1.8 of the samples studied, while the ranged values for concentrations ranged from (500-2400) ng /µl, specificity of the DNA extracted was confirmed using Zea mays specific gene responsible for production of Zein protein, a storage protein. Results shows that all the samples were positive for this gene, results of the investigation of sequence responsible for regulating gene expression for promoter P35s and T-nos terminator, should that 10 samples 13.9% of the total 72 samples studied are genetically modified and gave positive results for the amplification of PCR using primers specialized for each of the P35s and T-nos. The results indicated that (9 out of 47) represent 19.14% of the samples studied imported for the government institutions were genetically modified. Multiplex PCR technique used for the detection of two types of the targets at the same reaction to reduce the time and efforts. Multiplex PCR successfully applied for two combinations of either zein and P35s or zein and nos.

scholarly journals IUPAC Collaborative Trial Study of a Method To Detect Genetically Modified Soy Beans and Maize in Dried Powder

1999 ◽  
Vol 82 (4) ◽  
pp. 923-928 ◽  
Author(s):  
Markus Lipp ◽  
Peter Brodmann ◽  
Klaus Pietsch ◽  
Jean Pauwels ◽  
Elke Anklam ◽  
...  
Keyword(s):  
Genetically Modified Organisms ◽  
Genetically Modified ◽  
False Negative ◽  
35S Promoter ◽  
Lower Sensitivity ◽  
Correct Identification ◽  
Collaborative Trial ◽  
Negative Results ◽  
False Negative Results ◽  
Nos Terminator

Abstract This paper presents results of a collaborative trial study (IUPAC project No. 650/93/97) involving 29 laboratories in 13 countries applying a method for detecting genetically modified organisms (GMOs) in food. The method is based on using the polymerase chain reaction to determine the 35S promoter and the NOS terminator for detection of GMOs. Reference materials were produced that were derived from genetically modified soy beans and maize. Correct identification of samples containing 2% GMOs is achievable for both soy beans and maize. For samples containing 0.5% genetically modified soy beans, analysis of the 35S promoter resulted also in a 100% correct classification. However, 3 false-negative results (out of 105 samples analyzed) were reported for analysis of the NOS terminator, which is due to the lower sensitivity of this method. Because of the bigger genomic DNA of maize, the probability of encountering false-negative results for samples containing 0.5% GMOs is greater for maize than for soy beans. For blank samples (0% GMO), only 2 false-positive results for soy beans and one for maize were reported. These results appeared as very weak signals and were most probably due to contamination of laboratory equipment.

Multiplex, Construct-Specific, and Real-Time PCR-Based Analytical Methods for Bt Rice with cry1Ac Gene

2012 ◽  
Vol 95 (1) ◽  
pp. 186-194 ◽  
Author(s):  
Gurinder Jit Randhawa ◽  
Monika Singh
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Analytical Methods ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Pcr Assay ◽  
Cry1ac Gene ◽  
Camv 35S ◽  
Bt Rice ◽  
Pcr Assays

Abstract Qualitative and quantitative analytical methods based on PCR for Bacillus thuringiensis (Bt) rice hybrid, namely, MRP 5401 Bt expressing a modified version of the Bt cry1Ac gene, are reported here. Multiplex PCR assays were developed to target the cry1Ac transgene, Cauliflower mosaic virus (CaMV) 35S promoter, Agrobacterium tumefaciens nopaline synthase (nos) terminator, the neomycin phosphotransferase II (nptII) marker gene, and an endogenous α-tubulin (TubA) gene in Bt rice. The 3.178 kb region of inserted gene construct comprising the region of the CaMV 35S promoter and cry1Ac gene was amplified, and the construct integrity was confirmed by the nested PCR. The LOD for cry1Ac gene-specific simplex PCR was 0.01%, as established using Bt rice DNA dilutions with 100, 10, 1.0, 0.1, 0.05, 0.01, and 0.001% genetically modified trait. A real-time PCR assay was also developed to quantify the cry1Ac gene. The method performance of the reported real-time PCR assay was in line with the acceptance criteria of Codex Alimentarius Commission ALINORM 10/33/23, with LOD and LOQ values of 0.05%. The reliable PCR assays prior to commercial release of Bt rice would facilitate efficient regulatory compliance for identification of genetic trait, labeling requirements, and effective risk assessment and management. They could also address consumers' concerns and legal disputes that may arise.

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