Method for generating frangrance flavor through inactivation or reduction functional protein with betaine aldehyde dehydrogenase (BADH) activity
A protein and functional technology, applied in biochemical equipment and methods, redox enzymes, recombinant DNA technology, etc., can solve problems such as limited knowledge of biochemistry and genetics, molecular genetics that have not been described, and genes that have not been identified
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Embodiment 1
[0280] introduction
[0281] To identify the chromosomal location of genes encoding the rice aroma phenotype, simple sequence repeats (SSR) or microsatellite 2 and single nucleotide polymorphisms (SNPs) 3 Markers are mapped against segregating populations. Rice Genome Sequence 14 The availability of allows comparison of sequences of scented and non-scented genotypes. This allowed targeted resequencing of genes and genome sequences in the aroma genotype to the most probable portion of chromosome 8.
[0282] Materials and methods
[0283] plant material
[0284] In this study, 168 wild-grown F 2 Populations of individuals derived from Kyeema (Pelde / / Della / Kulu) (tall, jasmine-scented, scented, long-grain, Australian cultivar) and Gulfmont (Lebonnet / / CI9881 / PI 331581) (early maturing, semi-dwarf, non-scented, long-grain, American cultivar).
[0285] Following genetic mapping, candidate genes were identified in the region between the flanking markers of the published geno...
Embodiment 2
[0311] For any single analyst whose senses would become saturated, or often physically injured by the abrasion of the tongue from chewing hard grains, the ability to distinguish between scented and non-scented samples would increase with each successive analysis. decrease as the analysis proceeds.
[0312] Described below is the development of a PCR assay to genotype rice aroma.
[0313] plant material
[0314] All rice samples were provided by Yanco Agricultural Institute, NSW Agriculture. In addition to 168 wild-grown F 2 Individual populations were also validated using different collections of 14 scented and 74 non-scented varieties, where the F 2 Individuals derived from Kyeema (Pelde / / Della / Kulu) (tall, jasmine-scented, long-grained, Australian cultivar) and Gulfmont (Lebonnet / / CI9881 / PI 331581) (early, semi-dwarf, non-scented , long-grain, American cultivars).
[0315] genetic mapping
[0316] Genetic mapping was performed as described in Example 1.
[0317] Prime...
Embodiment 3
[0355] Materials and methods for generating transgenic wheat with BAD2 RNAi
[0356] Design RNAi inserts
[0357] The RNAi insert can be derived from the 5' end of the wheat BAD2 cDNA. This region shares 76.8% homology with the same region in the BAD1 gene homolog. In this example, RNAi inserts were designed such that transgenic plants exhibited specific BAD2 interference, but not BAD1 homolog interference (Figure 28).
[0358] PCR, genotyping and sequence analysis
[0359] Oligonucleotide primers are synthesized by methods known in the art. Use 0.2 μL Platinum(R) Taq DNA polymerase (Gibco BRL(R), 2 μL cDNA, 2.5 μL 10× buffer (Gibco BRL(R), 1 μL 50 mM MgCl 2 (Gibco BRL(R)), 1 μL of dNTPs (5 mM), 2.5 μL of each primer pair - (Table 1) [2 mM], in a total volume of 25 μL for PCR. PCR was performed using Perkin Elmer, Gene Amp PCR system9700. The cycle conditions are as follows: first denaturation at 94°C for 2 minutes, followed by 30 cycles at 94°C for 30 seconds, 58°C for ...
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