The Use of Next Generation Sequencing and Junction Sequence Analysis Bioinformatics to Achieve Molecular Characterization of Crops Improved Through Modern Biotechnology
| Autor: | Liang Guo, David K. Kovalic, Andre Silvanovich, Mingya Huang, Jeanna Groat, Jerry Hjelle, Qing Tian, Allen T. Christian, Lyle Ralston, Gary A. Bannon, Stephen R. Padgette, Nordine Cheikh, Yongpan Yan, Carl W. Garnaat |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Genetics
lcsh:QH426-470 Sequence analysis business.industry Locus (genetics) Plant Science Genetically modified crops lcsh:Plant culture Biology Bioinformatics Genome DNA sequencing law.invention Biotechnology lcsh:Genetics chemistry.chemical_compound chemistry law lcsh:SB1-1110 business Agronomy and Crop Science Polymerase chain reaction DNA Southern blot |
| Zdroj: | The Plant Genome, Vol 5, Iss 3, Pp 149-163 (2012) |
| ISSN: | 1940-3372 |
| Popis: | The assessment of genetically modifi ed (GM) crops for regulatory approval currently requires a detailed molecular characterization of the DNA sequence and integrity of the transgene locus. In addition, molecular characterization is a critical component of event selection and advancement during product development. Typically, molecular characterization has relied on Southern blot analysis to establish locus and copy number along with targeted sequencing of polymerase chain reaction products spanning any inserted DNA to complete the characterization process. Here we describe the use of next generation (NexGen) sequencing and junction sequence analysis bioinformatics in a new method for achieving full molecular characterization of a GM event without the need for Southern blot analysis. In this study, we examine a typical GM soybean [Glycine max (L.) Merr.] line and demonstrate that this new method provides molecular characterization equivalent to the current Southern blot-based method. We also examine an event containing in vivo DNA rearrangement of multiple transfer DNA inserts to demonstrate that the new method is effective at identifying complex cases. Next generation sequencing and bioinformatics offers certain advantages over current approaches, most notably the simplicity, effi ciency, and consistency of the method, and provides a viable alternative for effi ciently and robustly achieving molecular characterization of GM crops. M OLECULAR CHARACTERIZATION is a key step in the assessment of genetically modifi ed (GM) crops for regulatory approval. Current characterization methods are designed to accurately establish a number of important properties of the inserted DNA in the GM crop’s genome and this characterization is a key step in the production of improved GM crops. Th ese characterizations are required for selection of the events with the most favorable molecular profi le throughout research activities (Heck et al. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text