Quantitative analysis of phosphinothricin-N-acetyltransferase in genetically modified herbicide tolerant pepper by an enzyme-linked immunosorbent assay
- Authors
- Shim, Youn Young; Shin, Weon Sun; Moon, Gi Seong; Kim, Kyung Hwan
- Issue Date
- Apr-2007
- Publisher
- KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
- Keywords
- phosphinothricin-N-acetyltransferase; bar gene; GM pepper; sandwich ELISA
- Citation
- JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, v.17, no.4, pp.681 - 684
- Indexed
- SCIE
SCOPUS
KCI
- Journal Title
- JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
- Volume
- 17
- Number
- 4
- Start Page
- 681
- End Page
- 684
- URI
- https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/180269
- ISSN
- 1017-7825
- Abstract
- An immunoassay method was developed to quantitatively detect phosphinothricin-N-acetyltransferase (PAT) encoded by the Bialaphos resistance (bar) gene in genetically modified (GM) pepper. The histidine-tagged PAT was overexpressed in Escherichia coli M15 (pQE31-bar) and efficiently purified by Ni2+ affinity chromatography. A developed sandwich enzyme-linked immunosorbent assay (S-ELISA) method (detection limit: 0.01 mu g/ml) was 100-fold more sensitive than a competitive indirect ELISA (CI-ELISA) method or Western blot analysis in detecting the recombinant PAT. In real sample tests, PAT in genetically modified herbicide-tolerant (GMHT) peppers was successfully quantified [4.9 +/- 0.4 mu g/g of sample (n=6)] by the S-ELISA method. The S-ELISA method developed here could be applied to other GMHT crops and vegetables producing PAT.
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