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开发并验证一种整合的 DNA 游走策略，以检测表达 Cry 基因的转基因生物。

Development and validation of an integrated DNA walking strategy to detect GMO expressing cry genes.

机构信息

Scientific Institute of Public Health (WIV-ISP), Platform of Biotechnology and Bioinformatics (PBB) by: Sciensano, Transversal & Applied Genomics (TAG), J. Wytsmanstraat 14, 1050, Brussels, Belgium.

Scientific Institute of Public Health (WIV-ISP), Operational Direction Expertise, Service provisions & Customer relations by: Sciensano, Scientific Direction Expertise, Service provisions & Customer relations, J. Wytsmanstraat 14, 1050, Brussels, Belgium.

出版信息

BMC Biotechnol. 2018 Jun 27;18(1):40. doi: 10.1186/s12896-018-0446-x.

DOI:10.1186/s12896-018-0446-x
PMID:29945581
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6020286/
Abstract

BACKGROUND

Recently, an integrated DNA walking strategy has been proposed to prove the presence of GMO via the characterisation of sequences of interest, including their transgene flanking regions and the unnatural associations of elements in their transgenic cassettes. To this end, the p35S, tNOS and t35S pCAMBIA elements have been selected as key targets, allowing the coverage of most of GMO, EU authorized or not. In the present study, a bidirectional DNA walking method anchored on the CryAb/c genes is proposed with the aim to cover additional GMO and additional sequences of interest.

RESULTS

The performance of the proposed bidirectional DNA walking method anchored on the CryAb/c genes has been evaluated in a first time for its feasibility using several GM events possessing these CryAb/c genes. Afterwards, its sensitivity has been investigated through low concentrations of targets (as low as 20 HGE). In addition, to illustrate its applicability, the entire workflow has been tested on a sample mimicking food/feed matrices analysed in GMO routine analysis.

CONCLUSION

Given the successful assessment of its performance, the present bidirectional DNA walking method anchored on the CryAb/c genes can easily be implemented in GMO routine analysis by the enforcement laboratories and allows completing the entire DNA walking strategy in targeting an additional transgenic element frequently found in GMO.

摘要

背景

最近，提出了一种整合的 DNA 游走策略，通过对感兴趣的序列进行特征分析，包括其转基因侧翼区域和转基因盒中元素的非自然关联，来证明 GMO 的存在。为此，选择 p35S、tNOS 和 t35S pCAMBIA 元件作为关键靶标，允许覆盖大多数 GMO，无论是否获得欧盟授权。在本研究中，提出了一种基于 CryAb/c 基因的双向 DNA 游走方法，旨在覆盖额外的 GMO 和额外的感兴趣序列。

结果

首先，使用几种具有这些 CryAb/c 基因的 GM 事件评估了基于 CryAb/c 基因的拟议双向 DNA 游走方法的可行性。随后，通过低浓度的靶标（低至 20 HGE）研究了其灵敏度。此外，为了说明其适用性，在模拟 GMO 常规分析中分析的食品/饲料基质的样品上测试了整个工作流程。

结论

鉴于其性能评估的成功，基于 CryAb/c 基因的这种双向 DNA 游走方法可以由执行实验室轻松地应用于 GMO 常规分析中，并允许在靶向转基因元件中完成整个 DNA 游走策略，该元件经常在 GMO 中发现。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b23f/6020286/79e0a93de442/12896_2018_446_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b23f/6020286/79e0a93de442/12896_2018_446_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b23f/6020286/79e0a93de442/12896_2018_446_Fig1_HTML.jpg

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