scholarly journals Novel Method to Detect a Construct-Specific Sequence of the Acetolactate Synthase Gene in Genetically-Modified Flax CDC Triffid (FP967)

2010 ◽  
Vol 33 (3) ◽  
pp. 532-534 ◽  
Author(s):  
Kosuke Nakamura ◽  
Hiroshi Akiyama ◽  
Chihiro Yamada ◽  
Rie Satoh ◽  
Daiki Makiyama ◽  
...  
Keyword(s):  
Genetically Modified ◽  
Acetolactate Synthase ◽  
Specific Sequence ◽  
Novel Method

scholarly journals Evaluation of Modified PCR Quantitation of Genetically Modified Maize and Soybean Using Reference Molecules: Interlaboratory Study

2009 ◽  
Vol 92 (1) ◽  
pp. 223-233 ◽  
Author(s):  
Takashi Kodama ◽  
Hideo Kuribara ◽  
Yasutaka Minegishi ◽  
Satoshi Futo ◽  
Masatoshi Watai ◽  
...  
Keyword(s):  
Quantitative Methods ◽  
Conversion Factor ◽  
Recombinant Dna ◽  
Genetically Modified ◽  
Interlaboratory Study ◽  
Chain Reaction ◽  
Specific Sequence ◽  
Reference Plasmid ◽  
Repeatability And Reproducibility ◽  
Polymerase Chain

Abstract Real-time polymerase chain reaction (PCR)-based quantitative methods were previously developed and validated for genetically modified (GM) maize or soy. In this study, the quantification step of the validated methods was modified, and an interlaboratory study was conducted. The modification included the introduction of the PCR system SSIIb 3 instead of SSIIb 1 for the detection of the taxon-specific sequence of maize, as well as the adoption of colE1 as a carrier included in a reference plasmid solution as a replacement for salmon testis. The interlaboratory study was conducted with the ABI PRISM<sup/> 7700 and consisted of 2 separate stages: (1) the measurement of conversion factor (Cf) value, which is the ratio of recombinant DNA (r-DNA) sequence to taxon-specific sequence in each genuine GM seed, and (2) the quantification of blind samples. Additionally, Cf values of other instruments, such as the ABI PRISM 7900 and the ABI PRISM 7000, were measured in a multilaboratory trial. After outlier laboratories were eliminated, the repeatability and reproducibility for 5.0 samples were <15.8 and 20.6, respectively. The quantitation limits of these methods were 0.5 for Bt11, T25, and MON810, and 0.1 for GA21, Event176, and RR soy. The quantitation limits, trueness, and precision of the current modified methods were equivalent to those of the previous methods. Therefore, it was concluded that the modified methods would be a suitable replacement for the validated methods.

scholarly journals A Novel Method to Generate and Expand Clinical-Grade, Genetically Modified, Tumor-Infiltrating Lymphocytes

2017 ◽  
Vol 8 ◽  
Author(s):  
Marie-Andrée Forget ◽  
René J. Tavera ◽  
Cara Haymaker ◽  
Renjith Ramachandran ◽  
Shuti Malu ◽  
...  
Keyword(s):  
Genetically Modified ◽  
Tumor Infiltrating Lymphocytes ◽  
Clinical Grade ◽  
Novel Method ◽  
Infiltrating Lymphocytes

scholarly journals Increasing Yield of 2,3,5,6-Tetramethylpyrazine in Baijiu Through Saccharomyces cerevisiae Metabolic Engineering

2020 ◽  
Vol 11 ◽  
Author(s):  
Dan-Yao Cui ◽  
Ya-Nan Wei ◽  
Liang-Cai Lin ◽  
Shi-Jia Chen ◽  
Peng-Peng Feng ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Acetolactate Synthase ◽  
Fold Increase ◽  
Fermentation Performance ◽  
Strong Promoter ◽  
Rich Variety ◽  
Novel Method ◽  
Cerebrovascular Health ◽  
Decomposition Pathway ◽  
Butanediol Dehydrogenase

Baijiu is a traditional distilled beverage in China with a rich variety of aroma substances. 2,3,5,6-tetramethylpyrazine (TTMP) is an important component in Baijiu and has the function of promoting cardiovascular and cerebrovascular health. During the brewing of Baijiu, the microorganisms in jiuqu produce acetoin and then synthesize TTMP, but the yield of TTMP is very low. In this work, 2,3-butanediol dehydrogenase (BDH) coding gene BDH1 and another BDH2 gene were deleted or overexpressed to evaluate the effect on the content of acetoin and TTMP in Saccharomyces cerevisiae. The results showed that the acetoin synthesis of strain α5-D1B2 was significantly enhanced by disrupting BDH1 and overexpressing BDH2, leading to a 2.6-fold increase of TTMP production up to 10.55 mg/L. To further improve the production level of TTMP, the α-acetolactate synthase (ALS) of the pyruvate decomposition pathway was overexpressed to enhance the synthesis of diacetyl. However, replacing the promoter of the ILV2 gene with a strong promoter (PGK1p) to increase the expression level of the ILV2 gene did not result in further increased diacetyl, acetoin and TTMP production. Based on these evidences, we constructed the diploid strains AY-SB1 (ΔBDH1:loxP/ΔBDH1:loxP) and AY-SD1B2 (ΔBDH1:loxP-PGK1p-BDH2-PGK1t/ΔBDH1:loxP-PGK1p-BDH2-PGK1t) to ensure the fermentation performance of the strain is more stable in Baijiu brewing. The concentration of TTMP in AY-SB1 and AY-SD1B2 was 7.58 and 9.47 mg/L, respectively, which represented a 2.3- and 2.87-fold increase compared to the parental strain. This work provides an example for increasing TTMP production in S. cerevisiae by genetic engineering, and highlight a novel method to improve the quality and beneficial health attributes of Baijiu.

scholarly journals Subspecies-specific sequence detection for differentiation of Mycobacterium abscessus complex

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Alina Minias ◽  
Lidia Żukowska ◽  
Jakub Lach ◽  
Tomasz Jagielski ◽  
Dominik Strapagiel ◽  
...  
Keyword(s):  
Dna Hybridization ◽  
Mycobacterium Abscessus ◽  
Human Pathogens ◽  
Specific Sequence ◽  
Sequence Detection ◽  
Virtual Database ◽  
Mycobacterium Abscessus Complex ◽  
Etiologic Agents ◽  
Signature Sequences ◽  
Novel Method

Abstract Mycobacterium abscessus complex (MABC) is a taxonomic group of rapidly growing, nontuberculous mycobacteria that are found as etiologic agents of various types of infections. They are considered as emerging human pathogens. MABC consists of 3 subspecies—M. abscessus subsp. bolletti, M. abscessus subsp. massiliense and M. abscessus subsp. abscessus. Here we present a novel method for subspecies differentiation of M. abscessus named Subspecies-Specific Sequence Detection (SSSD). This method is based on the presence of signature sequences present within the genomes of each subspecies of MABC. We tested this method against a virtual database of 1505 genome sequences of MABC. Further, we detected signature sequences of MABC in 45 microbiological samples through DNA hybridization. SSSD showed high levels of sensitivity and specificity for differentiation of subspecies of MABC, comparable to those obtained by rpoB sequence typing.

Sign in / Sign up

Export Citation Format

Share Document