PubMed · 12802292
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2003-06-12. Debate, what debate?. https://doi.org/10.1038/423669b
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In many cases, multiple pathway enzymes need to be upregulated to produce a significant yield of a desired product. Technical advances in simultaneously manipulating multiple steps in plant metabolic pathways include the use of transcription factors, such as MYB12. By upregulating the genes of an entire pathway, these factors can greatly simplify multienzyme engineering. Furthermore, synthetic zinc-finger protein transcription factors can now be designed to target specific pathway enzymes, such as tocopherol methyltransferases. When multiple steps in a pathway are upregulated, previously unsuspected facets of the pathway might be revealed, such as the newly uncovered bifunctional substrate preference of the key regulatory enzyme in tocopherol (vitamin E) biosynthesis, homogentisate phytyltransferase. The engineering of desired traits, such as long-chain omega-3 polyunsaturated fatty acids, can require entirely new pathways to be introduced into a plant. Recent advances in genomics and gene expression technology have made this type of complex metabolic engineering highly feasible.
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Because the labeling of grains and feed- and foodstuffs is mandatory if the genetically modified organism (GMO) content exceeds a certain level of approved genetically modified varieties in many countries, there is a need for a rapid and useful method of GMO quantification in food samples. In this study, a rapid detection system was developed for Roundup Ready Soybean (RRS) quantification using a combination of a capillary-type real-time PCR system, a LightCycler real-time PCR system, and plasmid DNA as the reference standard. In addition, we showed for the first time that the plasmid and genomic DNA should be similar in the established detection system because the PCR efficiencies of using plasmid DNA and using genomic DNA were not significantly different. The conversion factor (Cf) to calculate RRS content (%) was further determined from the average value analyzed in three laboratories. The accuracy and reproducibility of this system for RRS quantification at a level of 5.0% were within a range from 4.46 to 5.07% for RRS content and within a range from 2.0% to 7.0% for the relative standard deviation (RSD) value, respectively. This system rapidly monitored the labeling system and had allowable levels of accuracy and precision.