제초제 내성 유전자 변형 옥수수 중 PAT단백질에 특이한 단크론성 항체의 생산과 특성 확인

다국어 초록 (Multilingual Abstract)

In this study, PAT protein of genetically modified maize was prepared from the recombinant E. coli strain BL21 (DE3), and mice were immunized with the recombinant PAT protein. After cell fusion and cloning, two hybridoma cells (PATmAb-7 and PATmAb-12) were chosen since the monoclonal antibodies (Mabs) produced by them were confirmed to be specific to PAT protein in the indirect enzyme-linked immunsorbent assay (ELISA) and western blot tests. There were no cross-reactions of either Mabs to other GM proteins or to the extracts of non-GM maize. The ELISA based on the PATmAb-7 can sensitively detect 0.3 ng/g PAT protein in corn. These results indicate that the developed Mabs can be used as bio-receptors in the development of immunosensors and biosensors for the rapid and simple detection of GM corn adulterated in foods.

목차 (Table of Contents)

• ABSTRACT
• Materials and Methods
• 실험재료 및 기기
• PAT 단백질의 확보
• 단크론성 항체 확보를 위한 hybridoma 세포주 개발
• PAT에 대한 단크론성 항체 생산 및 정제
• 생산된 PAT 단크론성 항체의 특성 확인
• Results and Discussion
• 재조합 PAT 단백질의 생산 및 정제
• 재조합 PAT 단백질의 항원성 확인 및 hybridoma 개발
• 단크론성 항체 정제 및 특성 확인
• 국문요약
• References

참고문헌 (Reference)

1 신원선, "유전자재조합 감자의 검정을 위한 DNA분리 및 PCR검출의 최적조건 탐색" 한국식품과학회 35 (35): 591-597, 2003

2 "https://www.foodsafetykorea.go.kr/portal/board/boardDetail.do/"

3 "http://www.fao.org/fileadmin/user_upload/gmfp/docs/SYHT0H2_report.pdf/"

4 "http://foodsafetykorea.go.kr/portal/board/board.do/"

5 Delaney, B., "Thirteen week rodent feeding study with grain from molecular stacked trait lepidopteran and coleopteran protected (DP-OO4114-3) maize" 53 : 417-427, 2013

6 Ford, C.S., "Spontaneous gene flow from rapeseed (Brassica napus) to wild Brassica oleracea" 273 : 3111-3115, 2006

7 Mckearn, T.J., "Selective suppression of humoral and cell-mediated immune responses to rat alloantigens by monoclonal antibodies produced by hybridoma cell lines" 11 : 932-935, 1979

8 Hooftman, D.A.P., "Seed bank dynamics govern persistence of Brassica hybrids in crop and natural habitats" 115 : 1-11, 2014

9 Zou, S., "Safety assessment of lepidopteran insect-protected transgenic rice with cry2A gene" 25 : 163-172, 2016

10 Keese, P., "Risks from GMOs due to horizontal gene transfer" 7 : 123-149, 2008

11 Cao, S.S., "Metabonomics study of transgenic Bacillus thuringiensis rice (T2A-1) meal in a 90-day dietary toxicity study in rats" 7 : 2304-2310, 2011

12 Duke, S.O., "Glyphosate effects on plant mineral nutrition, crop rhizosphere microbiota, and plant disease in glyphosate-resistant. Crops" 60 : 10375-10397, 2012

13 Bonny, S., "Genetically modied herbicide-tolerant crops, weeds, and herbicides: Overview and impact" 57 : 31-48, 2016

14 Chung, K.H., "Genetically Modified Organisms" 64-73, 2011

15 Xu, W.T., "Enzyme linked immunosorbent assay for PAT protein detection in genetically modified rape" 3 : 177-181, 2006

16 Towbin, H., "Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications" 76 : 4350-4354, 1979

17 George, C.W., "Effects of ammonium phosphate and sulphate on the pyrolysis and combustion of cellulose. USDA Forest Service INT-90" 1971

18 Shon, D.H., "Development of immunoassay system for the detection of genetically modified soybeans" 15 : 59-69, 2002

19 Kohler, G., "Continuous cultures of fused cells producing antibody of predefined specificity" 256 : 495-501, 1975

20 Nilufer, D., "Comparative study of three different methods for the determination of aflatoxins in tahini" 50 : 3375-3379, 2002

21 Laemmli, U.K., "Cleavage of structural proteins during the assembly of the head of bacteriophage T4" 15 : 680-685, 1970

22 Devos, Y., "Bt-maize event MON 88017 expressing Cry3Bb1 does not cause harm to non-target organisms" 21 : 1191-1214, 2012

23 Ford, C.S., "Assessing the value of imperfect biocontainment nationally: rapeseed in the United Kingdom as an exemplar" 205 : 1342-1349, 2015

24 Langone, J.J., "Aflatoxin B; specific antibodies and their use in radioimmunoassay" 56 : 591-595, 1976

25 Zhu, H.J., "A 90day safety assessment of genetically modified rice expressing Cry1Ab/1Ac protein using an aquatic animal model" 63 : 3627-3633, 2015