Detection of CaMV-35S Promoter and NOS Terminator in Genetically Modified Tomato Seed in Iraqi Markets

Abstract

The present study was focused to detect leading elements that control gene expression in genetically modified tomato by using conventional PCR technique.These common elements in all GM. plants are CaMV-35S promoter isolated from cauliflower mosaic virus and T-Nos terminator from the Agrobacterium tumefaciens. Seventy eight tomato genotypes were collected from Iraqiinstitutions and markets. The experiment was conducted in the Institute of Genetic Engineering/University of Baghdad/ Iraq and Directorate of Seeds Testing and Certification/Ministry of Agriculture/ Iraq. The tomato DNA samples were extracted manually by C- hexadecyl- Trimethyl-Ammonium-Bromide (CTAB) method. When measuring the optical density (OD) of the tomato samples, most purity values were found to be between (1.7-1.9).Two specific primers of CaMV-35S promoter, Nos terminator supplied by Canadian Alpha DNA Company, AccuPower®PCR Pre mix PCR supplied by Korean Bioneer Company and positive control (plasmid) supplied by Dr. ShathaAyidYousif/ Directorate of Agricultural Research/ Ministry of Science and Technology/ Iraq, were used in this study. Results showed that twenty four tomato genotypes were genetically modified. The primer specific of CaMV-35S promoter recorded a PCR product of ‎‎195 bp in 15 GM tomato and 13 GM tomato genotypes contain Nos terminator were a PCR product of ‎‎180 bp which as match with results of positive control (plasmid) which contains promoter and terminatorand that four tomato genotypes contain major components CaMV-35S promoter and Nos terminator together in the same sample.