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A real-time PCR-based assay was developed to measure the expression of the Cry1Ac genes in the Bacillus thuringiensis subsp. israelensis (Bti) genome by determining the abundance of the nptII gene. The soil microcosms were supplemented with 50, 500 and 1000 ng Bti DNA per gram of soil and were maintained for 7, 14, 28 and 42 days. By week 7, the abundance of Bti gene copies in soil was highest (6.3 x 10(5) copies/g of soil) when 1000 ng DNA was applied, but the abundance decreased with a decrease in the quantity of DNA. The efficiency of the Bti DNA incorporation into soil DNA was higher than that of kanamycin sulfate DNA. From 7 to 42 days, the abundance of the nptII gene decreased as the number of gene copies increased. The Bti DNA was not detected from 28 to 42 days, indicating that the Bti was not able to survive at the same time as the nptII gene. The number of Cry1Ac gene copies in soil was below the detection limit of the real-time PCR-based assay from 7 to 28 days, and the total number of copies did not change from 28 to 42 days. The abundance of the Cry1Ac gene was greatest in the root, followed by the stems, leaves, eggs and the soil. This assay can detect the Bti genes in
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