• Title/Summary/Keyword: molecular map

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Identification of QTLs Associated with Physiological Nitrogen Use Efficiency in Rice

  • Cho, Young-Il;Jiang, Wenzhu;Chin, Joong-Hyoun;Piao, Zhongze;Cho, Yong-Gu;McCouch, Susan R.;Koh, Hee-Jong
    • Molecules and Cells
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    • v.23 no.1
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    • pp.72-79
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    • 2007
  • Demand for low-input sustainable crop cultivation is increasing to meet the need for environment-friendly agriculture. Consequently, developing genotypes with high nutrient use efficiency is one of the major objectives of crop breeding programs. This study was conducted to identify QTLs for traits associated with physiological nitrogen use efficiency (PNUE). A recombinant inbred population (DT-RILs) between Dasanbyeo (a tongil type rice, derived from an indica ${\times}$ japonica cross and similar to indica in its genetic make-up) and TR22183 (a Chinese japonica variety) consisting of 166 $F_8$ lines was developed and used for mapping. A frame map of 1,409 cM containing 113 SSR and 103 STS markers with an average interval of 6.5 cM between adjacent marker loci was constructed using the DT-RILs. The RILs were cultivated in ordinary-N ($N-P_2O_5-K_2O=100-80-80kg/ha$) and low-N ($N-P_2O_5-K_2O=50-80-80kg/ha$) (100 kg/ha) conditions. PNUE was positively correlated with the harvest index and grain yield in both conditions. Twenty single QTLs (S-QTLs) and 58 pairs of epistatic loci (E-QTLs) were identified for the nitrogen concentration of grain, nitrogen concentration of straw, nitrogen content of shoot, harvest index, grain yield, straw yield and PNUE in both conditions. The phenotypic variance explained by these S-QTLs and E-QTLs ranged from 11.1 to 44.3% and from 16.0% to 63.6%, respectively. The total phenotypic variance explained by all the QTLs for each trait ranged from 35.8% to 71.3%, showing that the expression of PNUE and related characters depends signify- cantly upon genetic factors. Both S-QTLs and E-QTLs may be useful for marker-assisted selection (MAS) to develop higher PNUE genotypes.

Eco-toxicogenomics Research with Fish

  • Park, Kyeong-Seo;Kim, Han-Na;Gu, Man-Bock
    • Molecular & Cellular Toxicology
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    • v.1 no.1
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    • pp.17-25
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    • 2005
  • There are some critical drawbacks in the use of biomarkers for a global assessment of the toxicological impacts many chemicals and environmental pollutants have, primarily due to an individual biomarker's specificity for an explicit chemical or toxicant. In other words, the biomarker-based assessment methodology used to analyze toxicological effects lacks a high-throughput capability. Therefore, eco-toxicogenomics, or the study of toxicogenomics with organisms present within a given environmental locale, has recently been introduced with the advent of the so-called "-omics" era, which began with the creation of microarray technologies. Fish are comparable with humans in their toxicological responses and thus data from toxicogenomic studies performed with fish could be applied, with appropriate tools and implementation protocols, to the evaluation of environments where human or animal health is of concern. At present, there have been very active research streams for developing expression sequence tag (EST) databases (DBs) for zebra fish and rainbow trout. Even though few reports involve toxicogenomic studies with fish, a few groups have successfully fabricated and used cDNA microarrays or oligo DNA chips when studying the toxicological impacts of hypoxia or some toxicants with fish. Furthermore, it is strongly believed that this technology can also be implemented with non-model fish. With the standardization of DNA microarray technologies and ample progress in bioinformatics and proteomic technologies, data obtained from DNA microarray technologies offer not only multiple biomarker assays or an analysis of gene expression profiles, but also a means of elucidating gene networking, gene-gene relations, chemical-gene interactions, and chemical-chemical relationships. Accordingly, the ultimate target of eco-toxicogenomics should be to predict and map the pathways of stress propagation within an organism and to analyze stress networking.

Molecular Cloning of Chitinase Genes Family from Serratia marcescens

  • Song, Young-Hwan;Kweon, Oh-Gun
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.103-110
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    • 1993
  • Total genomic DNA library of Serratia marcescens was prepared by inserting Sau3AI partial digesting fragments(above 5 kb) into the dephosphorylated BamHl site of pUC19. In primary screening, two colonies were selected by observing the halo around E. coli transformants grown on the swollen colloidal chitin media. Secondary screening was performed by soaking two colonies with a few drops of 4-methylumbelleliferryl N-acetyl-$\beta$-D-glucocosaminide(4-MuNGlcNAc). As 4-MuNGlcNAc is a specific, fluorogenic substrate for chitinase, the positive clones produce light fluorescence by the exposure under the long wave U.V. light(360 nm). From genomic DNA library derived from pUC19, we have isolated two different chitinase clones, pCH1(11.0Kb) and pCH2(7.5Kb), which show completely different restriction map to each other. The cross-hybridization of pCH1EA and pCH2 have not revealed any hybridization signals to each other.

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Molecular Cloning of nifHD from Rhizobium sp. SNU003 (Rhizobium sp. SNU003의 nifHD 클로닝)

  • 강명수;안정선
    • Korean Journal of Microbiology
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    • v.31 no.2
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    • pp.123-128
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    • 1993
  • Genes for dinitrogenase reductase (nifH) and dinitogenase a subunit (nifD) were found to be located on 7.9 kb of EcoRI, 6.5 kb of Sail, 7.3 kb of HindlII and 4.4 kb of Pstl fragments of the genomic blot of Rhizobium sp. SNU003. a symbiotic strain from root nodule of Canavalia lineata. Nine recombinant phage nif-clones were selected from the genomic library constructed by using EMBL-3 BamHI arms of bacteriophage lambda. Among them. Rnif-6 had insert DNA of 15.3 kb. in which 7.6 kb of BamHI!SacI fragment contained nifHD region. Therefore, the 7.6 kb fragment was subcloned into pUC19 and partial restriction map was constructed. As the results, nifH and nifD were found to be located continuously on 4.5 kb of BamHI/BglIl in the genome of Rhizobium sp. SNU003 strain.

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Sequence Analysis, Molecular Cloning and Restriction Mapping of Mitochondreal Genome of Domesticated Silkworm, Bombyx mori (누에 미토콘드리아 유전체의 제한효소 지도작성, 클로닝 및 염기서열 분석)

  • 이진성;성승현;김용성;서동상
    • Journal of Sericultural and Entomological Science
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    • v.42 no.1
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    • pp.14-23
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    • 2000
  • The mitochondrial genome of domesticated silkworm (Bombyx mori) was mapped with five restriction endonucleases (BamHI, EcoRI, HindIII, PstI and XbaI), the entire genome was cloned with HindIII and EcoRI. From the end sequencing results of 5$^1$and 3$^1$region for full genome set of eleven mitochondrial clones, the seven mitochondrial genes (NADH dehydrogenase 6, ATPase 6, ATPase 8, tRN $A^{Lys}$, tRN $A^{Asp}$, tRN $A^{Thr}$ and tRN $A^{Phe}$ of mori were identified on the basis of their nucleotide sequence homology. The nucleotide composition of NADH dehydrogenase 6 was heavily biased towards adenine and thymine, which accounted for 87.76%. On basis of the sequence similarity with published tRNA genes from six insect species, the tRN $A^{Lys}$, tRN $A^{Asp}$ and tRN $A^{Thr}$ were showed stable canonical clover-leaf tRNA structures with acceptible anticodons. However, both the DHU and T$\psi$C arms of tRN $A^{Phe}$ could not form any stable stem-loop structure. The two overlapping gene pairs (tRN $A^{Lys}$ -tRN $A^{ASP}$ and ATPase8-ATPase6) were found from our sequencing results. The genes are encoded on the same strad. ATPase8 and ATPase6 overlaps (ATGATAA) which are a single example of overlapping events between abutted protein-coding genes are common, and there is evidence that the two proteins are transcribed from a single bicistronic message by initiation at 5$^1$terminal start site for ATPase8 and at an internal start site for ATPase6. Ultimately, this result will provide assistance in designing oligo-nucleotides for PCR amplification, and sequencing the specific mitochondrial genes for phylogenetics of geographic races, genetically improved silkworm strains and wild silkworm (mandarina) which is estimated as ancestal of domesticated silkworm.sticated silkworm.

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Comparison of Protein Profiles of Proso Millet (Panicum miliaceum) Seeds of Various Korean Cultivars

  • Roy, Swapan Kumar;Kwon, Soo-Jeong;Yu, Je-Hyeok;Sarker, Kabita;Cho, Seong-Woo;Moon, Young-Ja;Jung, Tae-Wook;Park, Cheol-Ho;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.62 no.1
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    • pp.40-50
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    • 2017
  • Seed storage proteins are used as carbon and nitrogen sources for the nutritional improvement of seeds. Since the composition of proteins from the Korean cultivars of proso millet is unknown, this study was conducted to obtain a reference map of millet seed proteins and identify the functional characteristics of the identified proteins. Proteins extracted from proso millet seeds of various cultivars were investigated using proteomic techniques such as 2-D electrophoresis coupled with mass fingerprinting; 1152 (differentially expressed) protein spots were detected on the 2-D gels. Among them, 26 reproducible protein spots were analyzed using matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry. Out of the 26 proteins, 2 proteins were upregulated in all the millet cultivars, while 13 proteins were upregulated and 11 proteins were downregulated in 2 cultivars. Abundance of most of the identified protein species associated with polysaccharide and starch metabolism, transcription, and pathogenesis was significantly enhanced, while that of other protein species involved in glycolysis, stress response, and transduction was severely reduced. Taken together, the results suggest that the differential expression of the proteins from the four millet cultivars may be cultivar-specific. By conducting a proteomic investigation of millet seeds from different cultivars, we sought to better understand the functional categorization of individual proteins on the basis of their molecular functions. We believe that the identified proteins may help in investigating genetic variations in millet cultivars.

Developmental Changes of Recessive Genes-mediated Cucumber mosaic virus (CMV) Resistance in Peppers (Capsicum annuum L.)

  • Min, Woong-Ki;Ryu, Jae-Hwang;Ahn, Su-Hyeon
    • Horticultural Science & Technology
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    • v.32 no.2
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    • pp.235-240
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    • 2014
  • Cucumber mosaic virus (CMV) is one of the most important viral diseases in pepper (Capsicum annuum L.), and several genes for resistance were reported in Capsicum spp. In Korea, a single dominant gene that is resistant to $CMV_{Fny}$ and $CMV_{P0}$ has been used for breeding. Recently, a new strain ($CMV_{P1}$) was reported that could infect cultivars resistant to both $CMV_{Fny}$ and $CMV_{P0}$. Therefore, breeding of more robust CMV-resistant cultivars is required. In this study, we surveyed the inheritance of $CMV_{P1}$ resistance and analyzed the location of the resistance loci. After $CMV_{P1}$ inoculation of various germplasms and breeding lines, one accession (ICPN18-8) showed no visual symptoms at 15 dpi (days post inoculation) but was susceptible after 45 dpi, and one resistant line (I7339) showed resistance until at 45 dpi. The latter line was used for tests of resistance inheritance. A total of 189 $F_2$ plants were examined, with 42 individuals showing resistance at 15 dpi and a phenotype segregation ratio close to 1:3 (resistant:susceptible plants). In a lateral ELISA test at 45 dpi, 11 plants showed resistance, and the segregation ratio was changed to 1:15. These results indicate that resistance in C. annuum 'I7339' is controlled by two different recessive genes; we named these resistance genes 'cmr3E' and 'cmr3L,' respectively. To locate these two resistant loci in the pepper linkage map, various RAPD, SSR, and STS markers were screened; only nine markers were grouped into one linkage group (LG). Only one RAPD primer (OPAT16) was distantly linked with cmr3E (22.3 cM) and cmr3L (20.7 cM). To develop more accurate markers for marker-assisted breeding, enriching for molecular markers spanning two loci will be required.

Investigation of Single Nucleotide Polymorphisms in Porcine Chromosome 2 Quantitative Trait Loci for Meat Quality Traits

  • Do, K.T.;Ha, Y.;Mote, B.E.;Rothschild, M.F.;Choi, B.H.;Lee, S.S.;Kim, T.H.;Cho, B.W.;Kim, K.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.2
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    • pp.155-160
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    • 2008
  • Several studies have reported quantitative trait loci (QTL) for meat quality on porcine chromosome 2 (http://www.animalgenome.org/QTLdb/pig.html). For application of the molecular genetic information to the pig industry through marker-assisted selection, single nucleotide polymorphism (SNP) markers were analyzed by comparative re-sequencing of polymerase chain reaction (PCR) products of 13 candidate genes with DNA from commercial pig breeds such as Berkshire, Yorkshire, Landrace, Duroc and Korean Native pig. A total of 34 SNPs were identified in 15 PCR products producing an average of one SNP in every 253 bp. PCR restriction fragment length polymorphism (RFLP) assays were developed for 11 SNPs and used to investigate allele frequencies in five commercial pig breeds in Korea. Eight of the SNPs appear to be fixed in at least one of the five pig breeds, which indicates that different selection among pig breeds might be applied to these SNPs. Polymorphisms detected in the PTH, CSF2 and FOLR genes were chosen to genotype a Berkshire-Yorkshire pig breed reference family for linkage and association analyses. Using linkage analysis, PTH and CSF2 loci were mapped to pig chromosome 2, while FOLR was mapped to pig chromosome 9. Association analyses between SNPs in the PTH, CSF2 and FOLR suggested that the CSF2 MboII polymorphism was significantly associated with several pork quality traits in the Berkshire and Yorkshire crossed F2 pigs. Our current findings provide useful SNP marker information to fine map QTL regions on pig chromosome 2 and to clarify the relevance of SNP and quantitative traits in commercial pig populations.

Barbigerone Inhibits Tumor Angiogenesis, Growth and Metastasis in Melanoma

  • Yang, Jian-Hong;Hu, Jia;Wan, Li;Chen, Li-Juan
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.167-174
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    • 2014
  • Tumor angiogenesis, growth and metastasis are three closely related processes. We therefore investigated the effects of barbigerone on all three in the B16F10 tumor model established in both zebrafish and mouse models, and explored underlying molecular mechanisms. In vitro, barbigerone inhibited B16F10 cell proliferation, survival, migration and invasion and suppressed human umbilical vascular endothelial cell migration, invasion and tube formation in concentration-dependent manners. In the transgenic zebrafish model, treatment with $10{\mu}M$ barbigerone remarkably inhibited angiogenesis and tumor-associated angiogenesis by reducing blood vessel development more than 90%. In vivo, barbigerone significantly suppressed angiogenesis as measured by H and E staining of matrigel plugs and CD31 staining of B16F10 melanoma tumors in C57BL/6 mice. Furthermore, it exhibited highly potent activity at inhibiting tumor growth and metastasis to the lung of B16F10 melanoma cells injected into C57BL/6 mice. Western blotting revealed that barbigerone inhibited phosphorylation of AKT, FAK and MAPK family members, including ERK, JNK, and p38 MAPKs, in B16F10 cells mainly through the MEK3/6/p38 MAPK signaling pathway. These findings suggested for the first time that barbigerone could inhibit tumor-angiogenesis, tumor growth and lung metastasis via downregulation of the MEK3/6/p38 MAPK signaling pathway. The findings support further investigation of barbigerone as a potential anti-cancer drug.

Functional PstI/RsaI Polymorphisms in the CYP2E1 Gene among South Indian Populations

  • Lakkakula, Saikrishna;Maram, Rajasekhar;Munirajan, Arasambattu Kannan;Pathapati, Ram Mohan;Visweswara, Subrahmanyam Bhattaram;Lakkakula, Bhaskar V.K.S.
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.179-182
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    • 2013
  • Human cytochrome P4502E1 (CYP2E1) is a well-conserved xenobiotic-metabolizing enzyme expressed in liver, kidney, nasal mucosa, brain, lung, and other tissues. CYP2E1 is inducible by ethanol, acetone, and other low-molecular weight substrates and may mediate development of chemically-mediated cancers. CYP2E1 polymorphisms alter the transcriptional activity of the gene. This study was conducted in order to investigate the allele frequency variation in different populations of Andhra Pradesh. Two hundred and twelve subjects belonging to six populations were studied. Genotype and allele frequency were assessed through TaqMan allelic discrimination (rs6413419) and polymerase chain reaction-sequencing (-1295G>C and -1055C>T) after DNA isolation from peripheral leukocytes. The data were compared with other available world populations. The SNP rs6413419 is monomorphic in the present study, -1295G>C and -1055C>T are less polymorphic and followed Hardy-Weinberg equilibrium in all the populations studied. The -1295G>C and -1055C>T frequencies were similar and acted as surrogates in all the populations. Analysis of HapMap populations data revealed no significant LD between these markers in all the populations. Low frequency of $CYP2E1^*c2$ could be useful in the understanding of south Indian population gene composition, alcohol metabolism, and alcoholic liver disease development. However, screening of additional populations and further association studies are necessary. The heterogeneity of Indian population as evidenced by the different distribution of $CYP2E1^*c2$ may help in understanding the population genetic and evolutionary aspects of this gene.