• Title/Summary/Keyword: Molecular Detection

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Highly catalysis Zinc MOF-loaded nanogold coupled with aptamer to assay trace carbendazim by SERS

  • Jinling Shi;Jingjing Li;Aihui Liang;Zhiliang Jiang
    • Advances in nano research
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    • v.14 no.4
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    • pp.313-327
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    • 2023
  • Zinc metal organic framework (MOFZn)-loaded goad nanoparticles (AuNPs) sol (Au@MOFZn), which was characterized by TEM, Mapping, FTIR, XRD, and molecular spectrum, was prepared conveniently by solvothermal method. The results indicated that Au@MOFZn had a very strong catalytic effect with the nanoreaction of AuNPs formation between sodium oxalate (SO) and HAuCl4. AuNPs in the new indicator reaction had a strong resonance Rayleigh scattering (RRS) signal at 370 nm. The indicator AuNPs generated by this reaction, which had the most intense surface enhanced Raman scattering (SERS) peak at 1621 cm -1. The new SERS/RRS indicator reaction in combination with specific aptamer (Apt) to fabricate a sensitive and selective Au@MOFZn catalytic amplification-aptamer SERS/RRS assay platform for carbendazim (CBZ), with SERS/RRS linear range of 0.025-0.5 ng/mL. The detection limit was 0.02 ng/mL. Similarly, this assay platform has been also utilized to detect oxytetracycline (OTC) and profenofos (PF).

Genetic Insights into Domestication Loci Associated with Awn Development in Rice

  • Ngoc Ha Luong;Sangshetty G. Balkunde;Kyu-Chan Shim;Cheryl Adeva;Hyun-Sook Lee;Hyun-Jung Kim;Sang-Nag Ahn
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.33-33
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    • 2022
  • Rice (Oryza sativa L.) is a widely studied domesticated model plant. Seed awning is an unfavorable trait during rice harvesting and processing. Hence, awn was one of the target characters selected during domestication. However, the genetic mechanisms underlying awn development in rice are not well understood. In this study, we analyzed the genes for awn development using a mapping population derived from a cross between the Korean indica cultivar 'Milyang23' and NIL4/9 (derived from a cross between 'Hwaseong' and O. minuta). Two quantitative trait loci (QTLs), qAwn4 and qAwn9 were mapped on chromosome 4 and 9, respectively, increased awn length in an additive manner. Through comparative sequencing analyses parental lines, LABA1 was determined as the causal gene underlying qAwn4. qAwn9 was mapped to a 199-kb physical region between markers RM24663 and RM24679. Within this interval, 27 annotated genes were identified, and five genes, including a basic leucine zipper transcription factor 76 (OsbZIP76), were considered candidate genes for qAwn9 based on their functional annotations and sequence variations. Haplotype analysis using the candidate genes revealed tropical japonica specific sequence variants in the qAwn9 region, which partly explains the non-detection of qAwn9 in previous studies that used progenies from interspecific crosses. This provides further evidence that OsbZIP76 is possibly a causal gene for qAwn9. The O. minuta qAwn9 allele was identified as a major QTL associated with awn development in rice, providing an important molecular target for basic genetic research and domestication studies. Our results lay the foundation for further cloning of the awn gene underlying qAwn9.

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Identification of Cell Type-Specific Effects of DNMT3A Mutations on Relapse in Acute Myeloid Leukemia

  • Seo-Gyeong Bae;Hyeoung-Joon Kim;Mi Yeon Kim;Dennis Dong Hwan Kim;So-I Shin;Jae-Sook Ahn;Jihwan Park
    • Molecules and Cells
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    • v.46 no.10
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    • pp.611-626
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    • 2023
  • Acute myeloid leukemia (AML) is a heterogeneous disease caused by distinctive mutations in individual patients; therefore, each patient may display different cell-type compositions. Although most patients with AML achieve complete remission (CR) through intensive chemotherapy, the likelihood of relapse remains high. Several studies have attempted to characterize the genetic and cellular heterogeneity of AML; however, our understanding of the cellular heterogeneity of AML remains limited. In this study, we performed single-cell RNA sequencing (scRNAseq) of bone marrow-derived mononuclear cells obtained from same patients at different AML stages (diagnosis, CR, and relapse). We found that hematopoietic stem cells (HSCs) at diagnosis were abnormal compared to normal HSCs. By improving the detection of the DNMT3A R882 mutation with targeted scRNAseq, we identified that DNMT3A-mutant cells that mainly remained were granulocyte-monocyte progenitors (GMPs) or lymphoid-primed multipotential progenitors (LMPPs) from CR to relapse and that DNMT3A-mutant cells have gene signatures related to AML and leukemic cells. Copy number variation analysis at the single-cell level indicated that the cell type that possesses DNMT3A mutations is an important factor in AML relapse and that GMP and LMPP cells can affect relapse in patients with AML. This study advances our understanding of the role of DNMT3A in AML relapse and our approach can be applied to predict treatment outcomes.

Twindemic Threats of Weeds Coinfected with Tomato Yellow Leaf Curl Virus and Tomato Spotted Wilt Virus as Viral Reservoirs in Tomato Greenhouses

  • Nattanong Bupi;Thuy Thi Bich Vo;Muhammad Amir Qureshi;Marjia Tabassum;Hyo-jin Im;Young-Jae Chung;Jae-Gee Ryu;Chang-seok Kim;Sukchan Lee
    • The Plant Pathology Journal
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    • v.40 no.3
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    • pp.310-321
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    • 2024
  • Tomato yellow leaf curl virus (TYLCV) and tomato spotted wilt virus (TSWV) are well-known examples of the begomovirus and orthotospovirus genera, respectively. These viruses cause significant economic damage to tomato crops worldwide. Weeds play an important role in the ongoing presence and spread of several plant viruses, such as TYLCV and TSWV, and are recognized as reservoirs for these infections. This work applies a comprehensive approach, encompassing field surveys and molecular techniques, to acquire an in-depth understanding of the interactions between viruses and their weed hosts. A total of 60 tomato samples exhibiting typical symptoms of TYLCV and TSWV were collected from a tomato greenhouse farm in Nonsan, South Korea. In addition, 130 samples of 16 different weed species in the immediate surroundings of the greenhouse were collected for viral detection. PCR and reverse transcription-PCR methodologies and specific primers for TYLCV and TSWV were used, which showed that 15 tomato samples were coinfected by both viruses. Interestingly, both viruses were also detected in perennial weeds, such as Rumex crispus, which highlights their function as viral reservoirs. Our study provides significant insights into the co-occurrence of TYLCV and TSWV in weed reservoirs, and their subsequent transmission under tomato greenhouse conditions. This project builds long-term strategies for integrated pest management to prevent and manage simultaneous virus outbreaks, known as twindemics, in agricultural systems.

Implementation of point-of-care platforms for rapid detection of porcine circovirus type 2

  • Chiao-Hsu Ke;Mao-Yuan Du;Wang-Ju Hsieh;Chiu-Chiao Lin;James Mingjuh Ting;Ming-Tang Chiou;Chao-Nan Lin
    • Journal of Veterinary Science
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    • v.25 no.2
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    • pp.28.1-28.11
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    • 2024
  • Background: Porcine circovirus type 2 (PCV2) infection is ubiquitous around the world. Diagnosis of the porcine circovirus-associated disease requires clinic-pathological elements together with the quantification of viral loads. Furthermore, given pig farms in regions lacking access to sufficient laboratory equipment, developing diagnostic devices with high accuracy, accessibility, and affordability is a necessity. Objectives: This study aims to investigate two newly developed diagnostic tools that may satisfy these criteria. Methods: We collected 250 specimens, including 170 PCV2-positive and 80 PCV2-negative samples. The standard diagnosis and cycle threshold (Ct) values were determined by quantitative polymerase chain reaction (qPCR). Then, two point-of-care (POC) diagnostic platforms, convective polymerase chain reaction (cPCR, qualitative assay: positive or negative results are shown) and EZtargex (quantitative assay: Ct values are shown), were examined and analyzed. Results: The sensitivity and specificity of cPCR were 88.23% and 100%, respectively; the sensitivity and specificity of EZtargex were 87.65% and 100%, respectively. These assays also showed excellent concordance compared with the qPCR assay (κ = 0.828 for cPCR and κ = 0.820 for EZtargex). The statistical analysis showed a great diagnostic power of the EZtargex assay to discriminate between samples with different levels of positivity. Conclusions: The two point-of-care diagnostic platforms are accurate, rapid, convenient and require little training for PCV2 diagnosis. These POC platforms can discriminate viral loads to predict the clinical status of the animals. The current study provided evidence that these diagnostics were applicable with high sensitivity and specificity in the diagnosis of PCV2 infection in the field.

Comparison of Results between Cytogenetic Technique and Molecular Genetic Technique in Colorectal Carcinoma Patients (대장암환자의 염색체 결실에서 세포유전학적 기법과 분자유전학적 기법의 결과 비교)

  • Park, Cheolin;Lee, Jae Sik
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.3
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    • pp.285-293
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    • 2017
  • Globally, 1.3 million people develop colon cancer every year, and 600,000 people die each year it. In Korea, colorectal carcinoma was associated with the highest death rate, accounting for 8,380 people, among solid cancers in 2015. Among the various methods for the diagnosis and study of colorectal carcinoma, the results obtained by cytogenetic and molecular genetic methods were compared. Detection rate was 47% in 18q, 40% in 17p, 27% in 22q, and 17% in 10q via CGH; detection rate was 57% in D18S59, 50% in D18S68, 50% in TP53CA, 47% in D18S6940% in D22S274, 37% in D22S283, 27% in D10S187, and 23% in D10S541 with LOH. Microsatellite marker matching rates were 100% in D22S274, 100% in D22S283, 100% in D10S186, 100% in D10S187, 100% in D10S541, 93% in D18S69, 93% in D18S68, 92% in TP53CA, and 89% in D18S59. The agreement rate between the two methods was 94.4% based on positive results using CGH. Based on the advantages of CGH, which was the ability to obtain information regarding the entire tumor genome at once, this experiment could identify the region with significant deletion using CGH and the more limited region LOH, with a completely different approach. LOH in the recurrent high-risk group, 18q21, was helpful in the selection of treatment modalities and in prognostic estimation as well as making the most appropriate decision for treatment. Therefore, it is suggested that LOH with surgical site tissues could be one of the treatment methods for recurrent high-risk group among patients with colorectal carcinoma.

Cyanobacteria and Secondary Metabolites (시아노박테리아의 이차대사물질에 대한 연구)

  • Kim, Gi-Eun;Kwon, Jong-Hee
    • KSBB Journal
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    • v.22 no.5
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    • pp.356-361
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    • 2007
  • Cyanobacteria are a very old group of prokaryotic organisms that produce very diverse secondary metabolites, especially non-ribosomal peptide and polyketide structures. Although some cyanobacteria produce lethal toxins such as microcystins and anatoxins, some may be useful either for development into commercial drugs or as biochemical tools. Detection of unknown secondary metabolites was carried in the present study by a screening of 98 cyanobacterial strains from Cyanobiotech GmbH in order to establish a screening process, isolate pure substances and determine their bioactivities. A degenerated polymerase chain reaction technique as molecular approaches has been used for general screening of NRPS gene and PKS gene in cyanobacteria. A putative PKS gene was detected by DKF/DKR primer in 38 strains (38.8%) and PCR amplicons resulted from a presence of NRPS gene were showed by MTF2/MTR2 primer in 30 strains (30.6%), respectively. A screening of interesting strains was performed by comparing PCR screening results with HPLC analyses of extracts. HPLC analysis for a detection of natural products was performed in extracts from biomass. 5 strains were screened for further scale-up processing. 7 pure substances were isolated from the scale-up cultures and tested for bioactivities under consideration to purity, amount and molecular weight of substances. One substance isolated from CBT 635 showed cytotoxic activity. This substance may be regarded as Microcystin LR.

Current status of Brassica A genome analysis (Brassica A genome의 최근 연구 동향)

  • Choi, Su-Ryun;Kwon, Soo-Jin
    • Journal of Plant Biotechnology
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    • v.39 no.1
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    • pp.33-48
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    • 2012
  • As a scientific curiosity to understand the structure and the function of crops and experimental efforts to apply it to plant breeding, genetic maps have been constructed in various crops. Especially, in the case of Brassica crop, genetic mapping has been accelerated since genetic information of model plant $Arabidopsis$ was available. As a result, the whole $B.$ $rapa$ genome (A genome) sequencing has recently been done. The genome sequences offer opportunities to develop molecular markers for genetic analysis in $Brassica$ crops. RFLP markers are widely used as the basis for genetic map construction, but detection system is inefficiency. The technical efficiency and analysis speed of the PCR-based markers become more preferable for many form of $Brassica$ genome study. The massive sequence informative markers such as SSR, SNP and InDels are also available to increase the density of markers for high-resolution genetic analysis. The high density maps are invaluable resources for QTLs analysis, marker assisted selection (MAS), map-based cloning and comparative analysis within $Brassica$ as well as related crop species. Additionally, the advents of new technology, next-generation technique, have served as a momentum for molecular breeding. Here we summarize genetic and genomic resources and suggest their applications for the molecular breeding in $Brassica$ crop.

Cloning and Characterization of a 5-Enolpyruvyl Shikimate 3-Phosphate Synthase (EPSPS) Gene from Korean Lawn Grass (Zoysia japonica) (들잔디 5-Enolpyruvyl Shikimate 3-Phosphate Synthase(EPSPS) 유전자 클로닝 및 특성)

  • Lee, Hye-Jung;Lee, Geung-Joo;Kim, Dong-Sub;Kim, Jin-Beak;Ku, Ja-Hyeong;Kang, Si-Yong
    • Horticultural Science & Technology
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    • v.28 no.4
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    • pp.648-655
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    • 2010
  • This study is the first comprehensive report on the molecular cloning, structural characterization, sequence comparison between wild and mutant types, copy number in the genome, expression features and activities of a gene encoding 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) in Korean lawn grass ($Zoysia$ $japonica$). The full length cDNA of the EPSPS from Korean lawn grass ($zj$EPSPS) obtained from a 3' and 5' RACE method was 1540 bp, containing a 1176 bp ORF, a 144 bp leader sequence (5' UTR) and a 220 bp 3' UTR, which was eventually decoded 391 amino acid residues with a molecular mass of 41.74 kDa. The Southern blot detection of the $zj$EPSPS showed that the gene exists as a single copy in the Korean lawn grass genome. Sequence comparison of the $zj$EPSPS gene demonstrated that the glyphosate-tolerant mutant (GT) having a Pro-53 to Ser substitution in the gene seems to have a preferred binding activity of the enzyme to phosphoenol pyruvate(PEP) over glyphosate, which allows the continuous synthesis of aromatic amino acids in the shikimate pathway. From the Northern blotting analysis, the $zj$EPSPS was found to be highly expressed, with continuous increase until 36 hours after 0.5% glyphosate treatment in both wild and mutant samples, but 1.5-fold higher EPSP synthase activity was observed in the tolerant mutant when exposed to the glyphosate treatment. The molecular information of the $zj$EPSPS gene obtained from this study needs to be further dissected to be more effectively applied to the development of gene-specific DNA markers and zoysiagrass cultivars; nevertheless, the glyphosate-tolerant mutant having the featured $zj$EPSPS gene can be provided to turfgrass managers for weed problems with timely adoptable management options.

MOLECULAR BIOLOGIC ANALYSIS OF FMR-1 GENE TRINUCLEOTIDE REPEATS IN AUTISTIC PATIENTS (자폐장애 환자에서 FMR-1 유전 삼염기 반복의 분자생물학적 분석)

  • Kwak, Ho-Soon;Chun, Hyo-Jin;Chang, Eun-Jin;Kim, Hee-Cheol;Kim, Jung-Bun;Park, Young-Nam;Jung, Chul-Ho
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.11 no.1
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    • pp.3-15
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    • 2000
  • Objectives:There has been a rapid expansion of studies aimed at elucidating the genetic basis of autistic disorder, especially it’ relationship to fragile-X syndrome. The detection of fragile X chromosome(Xq27.3) by cytogenetic analysis has revealed many difficulties in testing. Therefore, to explore the relationship between autistic disorder and fragile X syndrome, this study administered molecular biologic methods which examined an unstable CGG repeat within the fragile X mental retardation-1(FMR-1) gene. Methods:Ninety nine autistic children and eight normal control children were tested. The number of CGG repeats within FMR-1 gene was measured after amplification by PCR, and cytogenetic analysis was also carried out to detect fragile site Xq27.3. Southern blot hybridization, using StB12.3 and/or Pfxa3 probe, was done for the patients showing expansion of more than 50 CGG repeats (premutation). Results:All but two autistic patients had no expansion in CGG repeats by PCR and there was no significant statistical difference in number of CGG repeat in comparison with normal control. Two autistic patients, considered as premutation by PCR analysis, had no full mutation or premutation by Southern blot hybridization. All autistic children tested did not have any abnormal karyotype or fragile site Xq27.3. Conclusions:These results suggest that autistic patients may not have abnormality in FMR-1 gene or abnormal expansion in CGG repeat. In conclusion, fragile X syndrome may not be antecedent of autistic disorder.

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