Construction of a standard reference plasmid containing seven target genes for the detection of transgenic cotton

Wang, Xujing; Tang, Qiaoling; Dong, Lei; Dong, Yuchen; Su, Yueyan; Jia, Sun; Wang, Zhixing

doi:10.1016/j.plasmid.2014.05.005

Cited by 5 publications

(1 citation statement)

References 13 publications

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“…This situation is especially problematic in nonmodel systems. Therefore, it is important to consider this problem when faced with the need to genetically transform an organism at some level to test gene function (Wang et al 2014).…”

Section: Introductionmentioning

confidence: 99%

A plant transformation system designed for high throughput genomics inGossypium hirsutumto study root–organism interactions

Pant

McNeece

Sharma

et al. 2015

Journal of Plant Interactions

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The study of biological processes has been aided greatly by the development of procedures to identify the large numbers of associated genes. However, the ability to study the identified genes experimentally is often impeded by the absence of technologies to perform such functional analyses. Here, a nonaxenic plant transformation system has been developed in Gossypium hirsutum (cotton) for the study of genes associated with root functions and root-organism interactions. The plant transformation system is compatible with modern high throughput plant transformation goals and the processing of large numbers of genes intended for the study of root function in G. hirsutum.

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Section: Introductionmentioning

confidence: 99%

A plant transformation system designed for high throughput genomics inGossypium hirsutumto study root–organism interactions

Pant

McNeece

Sharma

et al. 2015

Journal of Plant Interactions

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Development and application of a multi-targeting reference plasmid as calibrator for analysis of five genetically modified soybean events

Cao

et al. 2015

Anal Bioanal Chem

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Reference materials are important in accurate analysis of genetically modified organism (GMO) contents in food/feeds, and development of novel reference plasmid is a new trend in the research of GMO reference materials. Herein, we constructed a novel multi-targeting plasmid, pSOY, which contained seven event-specific sequences of five GM soybeans (MON89788-5', A2704-12-3', A5547-127-3', DP356043-5', DP305423-3', A2704-12-5', and A5547-127-5') and sequence of soybean endogenous reference gene Lectin. We evaluated the specificity, limit of detection and quantification, and applicability of pSOY in both qualitative and quantitative PCR analyses. The limit of detection (LOD) was as low as 20 copies in qualitative PCR, and the limit of quantification (LOQ) in quantitative PCR was 10 copies. In quantitative real-time PCR analysis, the PCR efficiencies of all event-specific and Lectin assays were higher than 90%, and the squared regression coefficients (R(2)) were more than 0.999. The quantification bias varied from 0.21% to 19.29%, and the relative standard deviations were from 1.08% to 9.84% in simulated samples analysis. All the results demonstrated that the developed multi-targeting plasmid, pSOY, was a credible substitute of matrix reference materials, and could be used as a reliable reference calibrator in the identification and quantification of multiple GM soybean events.

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Detection of commercialized plant products derived from new genomic techniques (NGT) - Practical examples and current perspectives

Guertler¹,

Pallarz

Belter³

et al. 2023

Food Control

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Construction of a standard reference plasmid containing seven target genes for the detection of transgenic cotton

Cited by 5 publications

References 13 publications

A plant transformation system designed for high throughput genomics inGossypium hirsutumto study root–organism interactions

A plant transformation system designed for high throughput genomics inGossypium hirsutumto study root–organism interactions

Development and application of a multi-targeting reference plasmid as calibrator for analysis of five genetically modified soybean events

Detection of commercialized plant products derived from new genomic techniques (NGT) - Practical examples and current perspectives

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