• Title/Summary/Keyword: Candidate Gene

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Cloning, Expression, and Characterization of a Cold-Adapted and Surfactant-Stable Alginate Lyase from Marine Bacterium Agarivorans sp. L11

  • Li, Shangyong;Yang, Xuemei;Zhang, Lan;Yu, Wengong;Han, Feng
    • Journal of Microbiology and Biotechnology
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    • v.25 no.5
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    • pp.681-686
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    • 2015
  • The purpose of this study was to find a cold-adapted and surfactant-stable alginate lyase as a candidate for biotechnological and industrial applications. The gene for a new alginate lyase, AlyL1, from Agarivorans sp. L11 was cloned and expressed in Escherichia coli. The recombinant AlyL1 was most active at 40℃ (1,370 U/mg). It was a cold-adapted alginate lyase, which showed 54.5% and 72.1% of maximum activity at 15℃ and 20℃, respectively. AlyL1 was an alkaliphilic enzyme and most active at pH 8.6. In addition, it showed high stability in the presence of various surfactants at a high concentration (from 0.1% to 1% (w/v)). AlyL1 was an endo-type alginate lyase that degraded both polyM and polyG blocks, yielding disaccharides and trisaccharides as the main products. This is the first report of the cloning and functional expression of a cold-adapted and surfactant-stable alginate lyase. AlyL1 might be an interesting candidate for biotechnological and industrial applications.

Comparative Analysis of Repetitive Elements of Imprinting Genes Reveals Eleven Candidate Imprinting Genes in Cattle

  • Kim, HyoYoung;Kim, Heebal
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.6
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    • pp.893-899
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    • 2009
  • Few studies have reported the existence of imprinted genes in cattle compared to the human and mouse. Genomic imprinting is expressed in monoallelic form and it depends on a single parent-specific form of the allele. Comparative analysis of mammals other than the human is a valuable tool for explaining the genomic basis of imprinted genes. In this study, we investigated 34 common imprinted genes in the human and mouse as well as 35 known non-imprinted genes in the human. We found short interspersed nuclear elements (SINEs), long interspersed nuclear elements (LINEs), and long terminal repeats (LTRs) in imprinted (human and mouse) and control (cattle) genes. Pair-wise comparisons for the three species were conducted using SINEs, LINEs, and LTRs. We also calculated 95% confidence intervals of frequencies of repetitive sequences for the three species. As a result, most genes had a similar interval between species. We found 11 genes with conserved SINEs, LINEs, and LTRs in the human, mouse, and cattle. In conclusion, eleven genes (CALCR, Grb10, HTR2A, KCNK9, Kcnq1, MEST, OSBPL5, PPP1R9A, Sgce, SLC22A18, and UBE3A) were identified as candidate imprinted genes in cattle.

Possible Biomarker Gene for Radiation Workers in Hospital

  • Jin, Young-Woo;Jeong, Mee-Seon;Moon, Kien;Lee, Chee-Young;Bae, Sang-Woo;Choi, Soo-Yong;Lee, Yun-Sil
    • Molecular & Cellular Toxicology
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    • v.5 no.2
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    • pp.165-171
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    • 2009
  • Biomarkers indicating past exposure to radiation have not yet been entirely satisfactory. In this study, we validated several genes reported as radiation response genes, as biomarkers to detect past exposure to radiation in occupationally exposed workers, especially workers in the medical field. A total of 54 radiation workers in hospital were investigated for radiation exposure dose. Their average radiation dose of recent one year was 1.09 mSv ($\pm$1.63) with a 10.63 mSv ($\pm$12.91) cumulative dose. The results of the multiple regression analysis for the various variables indicate that the Hsc70 (P=0.0292) and ORAL (P=0.0045) may be candidate biomarkers for the recent 1 year radiation exposure in radiation workers, whereas AEN (P=0.0334) and PGAMI (P=0.0003) might be for cumulative exposure.

Expression profile identifies novel genes involved in neuronal differentiation

  • Kim, Jung-Hee;Lee, Tae-Young;Yoo, Kyung-Hyun;Lee, Hyo-Soo;Cho, Sun-A;Park, Jong-Hoon
    • BMB Reports
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    • v.41 no.2
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    • pp.146-152
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    • 2008
  • In the presence of NGF, PC12 cells extend neuronal processes, cease cell division, become electrically excitable, and undergo several biochemical changes that are detectable in developing sympathetic neurons. We investigated the expression pattern of the apoptosis-related genes at each stage of neuronal differentiation using a cDNA microarray containing 320 apoptosis-related rat genes. By comparing the expression patterns through time-series analysis, we identified candidate genes that appear to regulate neuronal differentiation. Among the candidate genes, HO2 was selected by real-time PCR and Western blot analysis. To identify the roles of selected genes in the stages of neuronal differentiation, transfection of HO2 siRNA in PC12 cells was performed. Down-regulation of HO2 expression causes a reduction in neuronal differentiation in PC12 cells. Our results suggest that the HO2 gene could be related to the regulation of neuronal differentiation levels.

Microbial Diversity in Swamp

  • Hong Soon Gyu;Lee Kang Hyun;Bae Kyung Sook
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2002.10a
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    • pp.90-93
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    • 2002
  • The revolution in molecular biology has given us greatly increased ability to obtain and to modify biological resources and to use them for the benefit of all humankind. The sequencing and the associated analysis of gene functions for a growing number of genomes will have an unprecedented effect on the uses of biological resources and the need for access to them. To investigate the diversity of microbial community in swamp, molecular systematic methods were applied. By amplified rDNA restriction analysis (ARDRA) and rDNA partial sequence analysis, $75\%$ of the isolates were known species. In case of uncultured analysis, almost all the selected clones were new species candidate. Especially archea and uncultured bacterial analyses, all clones were new taxon candidates. As for the eukaryotic diversity, several yeast form cultures were isolated from various samples of swamp. Among them, about $60\%$ of the isolates were easily identified. In case of a new species candidate, most strain were included in hymenomycetal yeasts.

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Beagle dogs parentage testing by using 22 ISAG microsatellite markers

  • Ji, Hye-jung;Kim, Eun-hee;Lee, Kyoung-kap;Kang, Tae-young;Lee, Joo-myoung;Shin, Hyoung-doo;Kim, Lyoung-hyo;Yun, Young-min
    • Korean Journal of Veterinary Research
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    • v.47 no.4
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    • pp.457-460
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    • 2007
  • The objective of the study was to establish routine parentage testing system in Beagle dogs using 22 ISAG (International Society for Animal Genetics) canine microsatellite markers (2005). Blood collections were obtained from a mother dog, 4 candidate father dogs and 3 offspring (n = 8). Genomic DNA samples were extracted from 8 Beagle dogs blood for PCR analysis. PCR products for the allele were analyzed by ABI 3130 DNA Sequencer and GeneScan (Ver 3.0) analysis and Genotyper (Ver. 2.1) software. The genetic relationship of mother and 3 offspring as well as one father dog among 4 candidate father dogs was confirmed by microsatellite allele analysis. The results of locus for amelogenin, which was designed for sexing, were matching with real gender among 8 Beagle dogs (female; 217/217 homozygosity, male; 179/217 heterozygosity). Twenty two ISAG microsatellite markers are useful the parentage test of Beagle dogs. In addition, amelogenin is an applicable marker to detecting real sex in dogs.

Development of Optimal Breeding Pigs Using DNA Marker Information

  • Kim, Sang-Wook;Roh, Jung-Gun;Cho, Yang-Il;Choi, Bong-Hwan;Kim, Tae-Hun;Kim, Jong-Joo;Kim, Kwan-Suk
    • Genomics & Informatics
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    • v.8 no.2
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    • pp.81-85
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    • 2010
  • The aim of the study was to investigate pig reference families, generated from Korean native pigs (KNP) that were crossed with Yorkshire (YS) breeds, which were used to evaluate genetic markers to select breeding animals with superior pork quality. A set of five candidate genes (PRKAG3, MC4R, CAST, ESR, and PRLR ) was analyzed for association with pork quality traits. PRKAG3 (I199V) SNP genotypes were significantly associated with muscle moisture, protein, and fat contents. The MC4R D298N polymorphism was significantly associated with meat tenderness and color traits. The CAST polymorphism was significantly associated with muscle moisture and crude protein traits. These three genes have been associated with pork quality traits in other pig populations, and some of our results are consistent with earlier studies. In addition, two reproductive candidate genes (ESR and PRLR ) did not have significant associations. These results suggest that further study is warranted to investigate and develop more DNA markers associated with pork quality in our KNP-crossed pig families.

12-Oxoeicosatetraenoic acid, a candidate signal for placenta separation, activates matrix metalloproteinase and induces apoptosis in bovine trophoblast cells

  • Hachiro Kamada
    • Animal Bioscience
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    • v.36 no.3
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    • pp.429-440
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    • 2023
  • Objective: 12-oxo-5Z,8Z,10E,14Z-eicosatetraenoic acid (12-KETE), a metabolite of arachidonic acid, is a strong candidate signal for placenta separation following calf discharge at delivery. In the present study, the effects of 12-KETE on bovine trophoblast cells were investigated to determine its function in the placentome at delivery. Methods: Bovine trophoblast cells derived from blastocysts were used. They were cocultured with or without fibroblasts derived from bovine placentome and/or bovine uterine epithelial cells. 12-KETE was added to the culture medium. Results: Bovine trophoblast cells contained binucleate cells and strongly expressed caudal type homeobox 2 (CDX-2) genes. Addition of 12-KETE to the trophoblast cell colony without feeder cells or that on a fibroblast monolayer induced rapid exfoliation of the colony. After 12-KETE addition, trophoblast cells emitted strong fluorescence caused by the degradation of dye-quenched collagen, indicating that 12-KETE activated matrix metalloproteinase of the trophoblast cells. Exfoliated cell colonies were stained with YOPRO-1, but not propidium iodide (PI). Moreover, DNA fragmentation and Bcl-2 associated X protein (Bax) gene (apoptosis stimulator) upregulation were observed in exfoliated cells, indicating that 12- KETE induced trophoblast cell apoptosis. These results were consistent with previous in vivo observations; however, even a lower concentration of 12-KETE activated trophoblast protease. Meanwhile, fibroblasts derived from the bovine placentome converted arachidonic acid to 12-KETE. Conclusion: These observations indicate that 12-KETE may serve as a signal for placenta separation at delivery.

Ranking Candidate Genes for the Biomarker Development in a Cancer Diagnostics

  • Kim, In-Young;Lee, Sun-Ho;Rha, Sun-Young;Kim, Byung-Soo
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2004.11a
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    • pp.272-278
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    • 2004
  • Recently, Pepe et al. (2003) employed the receiver operating characteristic (ROC) approach to rank candidate genes from a microarray experiment that can be used for the biomarker development with the ultimate purpose of the population screening of a cancer, In the cancer microarray experiment based on n patients the researcher often wants to compare the tumor tissue with the normal tissue within the same individual using a common reference RNA. This design is referred to as a reference design or an indirect design. Ideally, this experiment produces n pairs of microarray data, where each pair consists of two sets of microarray data resulting from reference versus normal tissue and reference versus tumor tissue hybridizations. However, for certain individuals either normal tissue or tumor tissue is not large enough for the experimenter to extract enough RNA for conducting the microarray experiment, hence there are missing values either in the normal or tumor tissue data. Practically, we have $n_1$ pairs of complete observations, $n_2$ 'normal only' and $n_3$ 'tumor only' data for the microarray experiment with n patients, where n=$n_1$+$n_2$+$n_3$. We refer to this data set as a mixed data set, as it contains a mix of fully observed and partially observed pair data. This mixed data set was actually observed in the microarray experiment based on human tissues, where human tissues were obtained during the surgical operations of cancer patients. Pepe et al. (2003) provide the rationale of using ROC approach based on two independent samples for ranking candidate gene instead of using t or Mann -Whitney statistics. We first modify ROC approach of ranking genes to a paired data set and further extend it to a mixed data set by taking a weighted average of two ROC values obtained by the paired data set and two independent data sets.

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The identification of novel regions for reproduction trait in Landrace and Large White pigs using a single step genome-wide association study

  • Suwannasing, Rattikan;Duangjinda, Monchai;Boonkum, Wuttigrai;Taharnklaew, Rutjawate;Tuangsithtanon, Komson
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.12
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    • pp.1852-1862
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    • 2018
  • Objective: The purpose of this study was to investigate a single step genome-wide association study (ssGWAS) for identifying genomic regions affecting reproductive traits in Landrace and Large White pigs. Methods: The traits included the number of pigs weaned per sow per year (PWSY), the number of litters per sow per year (LSY), pigs weaned per litters (PWL), born alive per litters (BAL), non-productive day (NPD) and wean to conception interval per litters (W2CL). A total of 321 animals (140 Landrace and 181 Large White pigs) were genotyped with the Illumina Porcine SNP 60k BeadChip, containing 61,177 single nucleotide polymorphisms (SNPs), while multiple traits single-step genomic BLUP method was used to calculate variances of 5 SNP windows for 11,048 Landrace and 13,985 Large White data records. Results: The outcome of ssGWAS on the reproductive traits identified twenty-five and twenty-two SNPs associated with reproductive traits in Landrace and Large White, respectively. Three known genes were identified to be candidate genes in Landrace pigs including retinol binding protein 7, and ubiquitination factor E4B genes for PWL, BAL, W2CL, and PWSY and one gene, solute carrier organic anion transporter family member 6A1, for LSY and NPD. Meanwhile, five genes were identified to be candidate genes in Large White, two of which, aldehyde dehydrogenase 1 family member A3 and leucine rich repeat kinase 1, associated with all of six reproduction traits and three genes; retrotransposon Gag like 4, transient receptor potential cation channel subfamily C member 5, and LHFPL tetraspan subfamily member 1 for five traits except W2CL. Conclusion: The genomic regions identified in this study provided a start-up point for marker assisted selection and estimating genomic breeding values for improving reproductive traits in commercial pig populations.