QUANTITATIVE DETECTION OF CAMV- 35S PROMOTER AND T-NOS TERMINATOR IN GENETIC MODIFIED TOMATO FROM IRAQI MARKETS

Abstract

Aim of this research was quantitative detection of Increase and decrease in copy number of CaMV-35S promoter and Nos terminator in genetic modified tomato by using Real-time PCR. Twenty four samples of genetic modified tomato seeds were isolated from 84 tomato samples collected from Iraqi markets during the period from December 2016 to January 2017. The experiences were conducted in the Institute of Genetic Engineering, University of Baghdad. Go Taq®qPCR master mix kit supplied by USA Promega Company, three specific primers to CaMV-35S promoter, T-Nos terminator and β-actin housekeeping gene supplied by Canadian Alpha Company were used. To quantitative detection of increase and decrease in copy number of GM tomato samples contain CaMV-35S promoter and T-Nos terminator, comparing with the β-actin (houskeeping gene) using Multiple of Median (MoM) equation. The results showed that the lowest recorded of Ct value was (27.88) for CaMV-35S promoter gave an increase in copy number (1.1766) above the normal limit, while highest recorded of Ct value was (32.67) gave an increase in copy number (1.0350) above the normal limit. The lowest recorded of Ct value was (27.35) for T-Nos terminator gave an increase in copy number (1.1600) above the normal limit, whereas highest recorded of Ct value was (32.82) gave a decrease in copy number (0.9920) under the normal limit.