NDLI: Event-specific qualitative and quantitative polymerase chain reaction methods for detection of insect-resistant genetically modified Chinese cabbage based on the 3′-junction of the insertion site

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Event-specific qualitative and quantitative polymerase chain reaction methods for detection of insect-resistant genetically modified Chinese cabbage based on the 3′-junction of the insertion site

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Journal of the Korean Society for Applied Biological Chemistry : Volume 55, Issue 2, April 2012

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Event-specific qualitative and quantitative polymerase chain reaction methods for detection of insect-resistant genetically modified Chinese cabbage based on the 3′-junction of the insertion site

Content Provider	Springer Nature Link
Author	Lim, Sun Hyung Lee, Jin Hyoung Suh, Seok Cheol Lim, Myung Ho Cho, Hyun Suk Kweon, Soon Jong Ahn, Hong Il Shin, Kong Sik Woo, Hee Jong
Copyright Year	2012
Abstract	Transgenic Chinese cabbage 416-3 was developed in Korea by a transformation event involving modified insectresistant gene (cry1Ac1). To monitor unintended release of genetically modified (GM) Chinese cabbage in the future, as well as to meet GM-labeling requirements, the development of a reliable method for detection of GM cabbage is requisite. To develop qualitative and quantitative polymerase chain reaction methods for the insect-resistant GM Chinese cabbage, a cytosolic glutathione reductase (BcgGR1) gene was used as the endogenous reference gene. Primer pairs CGR-1/-2, amplifying the Chinese cabbage endogenous gene, yielded an expected amplicon of 121 bp, whereas no amplified product was observed when DNA samples from seven non-cabbage plants were used as templates. The event-specific primer pairs amplifying the junction site between the endogenous genome sequence and the transferred DNA of GM event 416-3, produced amplicons of desired size by qualitative polymerase chain reaction (PCR) assay. An eventspecific quantitative PCR detection method was established using a TaqMan probe and a standard plasmid as a reference molecule, which contained both endogenous and event-specific sequences. For the validation of this method, three different compositions of w/w mixed samples (containing transgenic DNA at 5, 1, and 0.5% of total DNA in the control samples) were quantified. The precision, expressed as standard deviation (SD) and relative standard deviations (RSD), deviated by 0.03–0.26% and 4.75–8.06%, respectively. These results clearly demonstrate that the PCR methods developed herein can be used for event-specific qualitative and quantitative testings of insect-resistant GM Chinese cabbage.
Starting Page	367
Ending Page	375
Page Count	9
File Format	PDF
ISSN	17382203
Journal	Journal of the Korean Society for Applied Biological Chemistry
Volume Number	55
Issue Number	3
e-ISSN	2234344X
Language	English
Publisher	The Korean Society for Applied Biological Chemistry
Publisher Date	2012-06-30
Publisher Place	Seoul
Access Restriction	Subscribed
Subject Keyword	event-specific polymerase chain reaction (PCR) genetically modified organism detection reference molecule genetically modified Chinese cabbage Applied Microbiology Biological Techniques real-time PCR Bioorganic Chemistry
Content Type	Text
Resource Type	Article
Subject	Organic Chemistry Biochemistry, Genetics and Molecular Biology

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