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Title: A HISTOCHEMICAL METHOD USING A SUBSTRATE OF BETA-GLUCURONIDASE FOR DETECTION OF GENETICALLY MODIFIED PAPAYA

Author
item WAKUI, C. - NTL INST OF HLTH SCI
item AKIYAMA, H. - NTL INST OF HLTH SCI
item WATANABE, T. - NTL INST OF HLTH SCI
item Fitch, Maureen
item UCHIKAWA, S. - CIIF KOBE STATION
item KI, M. - OSAKA CITY INST PHES
item TAKAHASHI, K. - SAITAMA INST PUB HLTH
item CHIBA, R. - SHOWA PHARM UNIV
item FUJII, . - YOKOHAMA QURNTN STATION
item HINO, A. - NTL FOOD RES INST
item MAITANI, T. - NTL INST HLTH SCI

Submitted to: Trade Journal Publication
Publication Type: Trade Journal
Publication Acceptance Date: 10/15/2003
Publication Date: 2/1/2004
Citation: Wakui, C., Akiyama, H., Watanabe, T., Fitch, M.M., Uchikawa, S., Ki, M., Takahashi, K., Chiba, R., Fujii, .., Hino, A., Maitani, T. 2004. A histochemical method using a substrate of beta-glucuronidase for detection of genetically modified papaya. Journal of Food Hygenics Society of Japan. 45 (1): 19-24.

Interpretive Summary: Abstract only.

Technical Abstract: A histochemical assay for detecting genetically modified (GM) papaya (derived from Line 55-1) is described. GM papayas, currently undergoing a safety assessment in Japan, was developed using a construct that included a ß-glucuronidase (GUS) reporter gene linked to a virus coat protein (CP) gene. Histochemical assay was used a visualize the blue GUS reaction product from transgenic seed embryos. Twelve embryos per fruit were extracted from the papaya seeds using a surgical knife. The embryos were incubated with the substrate 5-bromo-4-chloro-3-indolyl- ß-D-glucuronide (X-Gluc) in a 96-well microtiter plate for 10-15 hours at 37oC. Seventy-five percent of GM papaya embryos should turn blue theoretically. The histochemical assay results were completely consistent with those from a qualitative polymerase chain reaction (PCR) method developed by this laboratory. Furthermore, the method was validated in a five-laboratory study. The method for detection of GM papaya is rapid and simple, and does not require use of specialized equipment.