Influence of Different Processing Treatments on the Detectability of Nucleic Acid and Protein Targets in Transgenic Soybean Meal.

Tian, Fang; Guan, Qingfeng; Wang, Xiumin; Teng, Da; Wang, Jianhua

Influences of dry heating, wet heating, and extrusion on the degradation of DNA and protein from transgenic soybean meal (TSM) were analyzed using qualitative PCR, quantitative real-time PCR (qPCR), indirect enzyme-linked immunosorbent assay (iELISA), and Western blot. The 414-bp Lectin gene was thoroughly degraded after dry heating between 75 and 90 °C for 30 min, wet heating, and extrusion at 165 °C with 39 % moisture content. The 483-bp CP4-EPSPS gene was not detected after dry heating, wet heating, and extrusion at 120 °C with 39 % moisture content. The degradation ratios of both Lectin and CP4-EPSPS genes increased from 0.4 to 92.1 % and 6.1 to 84.0 % as temperatures rose from 90 to 165 °C. iELISA results of the extruded TSM showed that the CP4-EPSPS protein content was reduced from 4.19 to 0.54 % as temperatures rose from 90 to 150 °C and was not detectable at 165 °C. Western blot results also showed the degradation of CP4-CPSPS protein after extrusion. Our results showed that temperature played an essential role in DNA and protein degradation, and the content of genetically modified organism (GMO) products may be changed after processing and could not reflect the initial content of the products.

SOYBEAN meal; ENZYME-linked immunosorbent assay; LECTIN genetics; WESTERN immunoblotting; TRANSGENIC organisms; NUCLEIC acid isolation methods

Applied Biochemistry & Biotechnology, 2014, Vol 172, Issue 7, p3686

0273-2289

Academic Journal

10.1007/s12010-014-0760-2