• Title/Summary/Keyword: Gene Mapping

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Full-length cDNA, Expression Pattern and Association Analysis of the Porcine FHL3 Gene

  • Zuo, Bo;Xiong, YuanZhu;Yang, Hua;Wang, Jun
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.10
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    • pp.1473-1477
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    • 2007
  • Four-and-a-half LIM-only protein 3 (FHL3) is a member of the LIM protein superfamily and can participate in mediating protein-protein interaction by binding one another through their LIM domains. In this study, the 5'- and 3'- cDNA ends were characterized by RACE (Rapid Amplification of the cDNA Ends) methodology in combination with in silico cloning based on the partial cDNA sequence obtained. Bioinformatics analysis showed FHL3 protein contained four LIM domains and four LIM zinc-binding domains. In silico mapping assigned this gene to the gene cluster MTF1-INPP5B-SF3A3-FHL3-CGI-94 on pig chromosome 6 where several QTL affecting intramuscular fat and eye muscle area had previously been identified. Transcription of the FHL3 gene was detected in spleen, liver, kidney, small intestine, skeletal muscle, fat and stomach, with the greatest expression in skeletal muscle. The A/G polymorphism in exon II was significantly associated with birth weight, average daily gain before weaning, drip loss rate, water holding capacity and intramuscular fat in a Landrace-derived pig population. Together, the present study provided the useful information for further studies to determine the roles of FHL3 gene in the regulation of skeletal muscle cell growth and differentiation in pigs.

Quantitative Trait Loci Mapping for Porcine Backfat Thickness

  • Wu, X.L.;Lee, C.;Jiang, J.;Peng, Y.L.;Yan, H.F.;Yang, S.L.;Xiao, B.N.;Liu, X.C.;Shi, Q.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.932-937
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    • 2002
  • A partial genome scan using porcine microsatellites was carried out to detect quantitative trait loci (QTL) for backfat thickness (BFT) in a pig reference population. This population carried QTL on chromosomes 1, 13 and 18. The QTL on chromosome 1 was located between marker loci S0113 and SW1301. The QTL corresponded to very low density lipoprotein receptor gene (VLDLR) in location and in biological effects, suggesting that VLDLR might be a candidate gene. The QTL found on chromosome 13 was found between marker loci SWR1941 and SW864, but significance for the marker-trait association was inconsistent by using data with different generations. The QTL on chromosome 18 was discovered between markers S0062 and S0117, and it was in proximity of the regions where IGFBP3 and GHRHR were located. The porcine obese gene might be also a candidate gene for the QTL on chromosome 18. In order to understand genetic architecture of BFT better, fine mapping and positional comparative candidate gene analyses are necessary.

Evaluation of a New Fine-mapping Method Exploiting Linkage Disequilibrium: a Case Study Analysing a QTL with Major Effect on Milk Composition on Bovine Chromosome 14

  • Kim, JongJoo;Georges, Michel
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.9
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    • pp.1250-1256
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    • 2002
  • A novel fine-mapping method exploiting linkage disequilibrium (LD) was applied to better refine the quantitative trait loci (QTL) positions for milk production traits on bovine chromosome 14 in the pedigree comprising 22 paternal half-sib families of a Black-and-White Holstein-Friesian grand-daughter design in the Netherlands for a total of 1,034 sons. The chromosome map was constructed with the 31 genetic markers spanning 90 Kosambi cM with the average inter-marker distance of 3.5 cM. The linkage analyses, in which the effects of sire QTL alleles were assumed random and the random factor of the QTL allelic effects was incorporated into the Animal Model, found the QTL for milk, fat, and protein yield and fat and protein % with the Lod scores of 10.9, 2.3, 6.0, 25.4 and 3.2, respectively. The joint analyses including LD information by use of multi-marker haplotypes highly increased the evidence of the QTL (Lod scores were 25.1, 20.9, 11.0, 85.7 and 17.4 for the corresponding traits, respectively). The joint analyses including DGAT markers in the defined haplotypes again increased the QTL evidence and the most likely QTL positions for the five traits coincided with the position of the DGAT gene, supporting the hypothesis of the direct causal involvement of the DGAT gene. This study strongly indicates that the exploitation of LD information will allow additional gains of power and precision in finding and localising QTL of interest in livestock species, on the condition of high marker density around the QTL region.

Isolation and Linkage Mapping of Coding Sequences from Chicken Cosmids by Exon Trapping

  • Mannen, H.;Dote, Y.;Uratsuji, H.;Yoshizawa, K.;Okamoto, S.;Tsuji, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.3
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    • pp.309-312
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    • 2004
  • We performed exon trapping in order to locate functional genes on chicken chromosomes (GGA) and to identify functional gene sequences from chicken cosmids. Sequence analysis of 100 clones revealed 17 putative exons, five of which were identified with known sequences in a gene database search: thymopoietin beta (TMPO), U5 snRNP-specific 40 kDa protein (HPRP8BP), dihydropyridine receptor alpha 1 subunit (CACNL1A3), cystein string protein (CPS) and C15orf4. We attempted to map the genes to chicken chromosomes by using FISH and linkage analysis. The chromosomal localizations were GGA1 (TMPO), GGA10 (C15orf4), GGA23 (HPRP8BP) and GGA28 (CPS) by FISH and linkage analysis, while that of CACNL1A3 was predicted to be on a microchromosome by FISH but not by linkage analysis. Comparative mapping analyses between chickens and humans for the genes revealed both known and new synteny. The syntenic conservation between GGA1 and human chromosome (HSA) 12q23 (TMPO) and between GGA10 and HSA15q25 (C15orf4), were consistent with a recent publication, while two new syntenies were observed between GGA28 and HSA20q13.3 in CPS and between GGA23 and HSA1p34-35 in HPRP8BP. The information of presently mapped genes can contribute as anchor markers based on functional genes and the construction of a comparative map.

QTL Mapping for 6-Year-Old Growths of a Single Open-Pollinated Half-Sib Family of a Selected Clone 7-1037 in Loblolly Pine(Pinus taeda) and Average Effect of QTL Allele Substitution (테다소나무 7-1037 클론의 단일 반형매 풍매가계 6년생 생장에 대한 QTL mapping과 QTL 대립유전자 치환의 평균효과)

  • Kim, Yong-Yul;Lee, Bong-Choon;O'Malley, David M.
    • Journal of Korean Society of Forest Science
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    • v.95 no.4
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    • pp.483-494
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    • 2006
  • We conducted QTL mapping for 6-year growths of open-pollinated half-sib progenies from a selected clone 7-1037 in Pinus taeda. With an AFLP marker analysis on haploid DNA samples from the megagametophytes of the open-pollinated seeds, we constructed 20 framework maps spanning a total of 1,869 cM in total length and 18.5 cM in an average interval length between markers. Composite interval mapping reveals that one QTL explains 5.9% of the total phenotypic variation of height, and three QTLs account for 3.9~5.6% of the variation of diameter at breast height (DBH). There are no correlations between the QTLs. The genetic effects of the QTLs are 39.6 cm in height and 7.20~9.41 mm in DBH, respectively, The average effects of gene substitution of the markers closely linked with the QTLs are 44.3 cm in height and 8.38~11.81 m in DBH. Under an assumption that the within-family heritability for the growth traits of loblolly pine is less than 0.2, the QTLs account for 26.8% of the additive genetic variance of the progenies. In terms of relative selection efficiency, the individual selection based on QTL markers could be 5 times as high as phenotypic selection. The results in this study indicate that the QTL mapping method with open-pollinated half-sib family could be more practical and applicable to the conventional seed orchard-based selection work than other mapping methods with a single full-sib family, in particular from the viewpoint that it can provide crucial information for within-family individual selection such as breeding value.

Molecular Cloning and Expression of the Acetyl Xylan Esterase Gene(estII) of Bacillus Stearothermophilus in Escherichia coli (Bacillus stearothermophilus Acetyl Exterase 유전자(estII)의 클로닝과 Escherichia coli에서의 발현)

  • Kim, Hee-Sun;Eom, Soo-Jung;Cho, Ssang-Goo;Choi, Yong-Jin
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.599-606
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    • 1994
  • Bacillus stearothermomophilus, a strong xylan degrader, was confirmed to express multiple esterase activities in addition to the major xylanolytic enzymes. One of the genes encoding the esterases was isolated from the genomic library of B. stearothermophilus constructed with EcoRl restriction endonuclease and pBR322 plasmid. Three recombinant plasmids showing the tributyrin degrading activity were selected from approximately 7, 000 E. coli HB101 transformants, and were found to have the same insert of a 3.2 kb DNA fragment. Restriction mapping and hybridization studies revealed that the gene(estII) on the hybrid plasmid (pKMG7) had originated from the B. stearothermophilus chromosome, and was distinct from the estl, another esterase gene of B. stearothermophilus isolated in the previous work. The E. coli cells harboring pKMG7 produced an acetylxylan esterase that exibited similar substrate specificity to the esterase encoded by the estI gene.

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Cloning of Isopenicillin N Synthase Gene from Lysobacter lactamgenus

  • Ryu, Jae-Kook;Nam, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.7 no.6
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    • pp.373-377
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    • 1997
  • The gene for isopenicillin N synthase (cyclase; IPNS) was cloned from Lysobacter lactamgenus using DNA probe amplified with primers based on the consensus sequences of isopenicillin N synthase genes of other ${\beta}$-lactam-producing microorganisms. The genomic library of L. lactamgenus using pUC18 plasmid cloned at the SacI site were screened with the PCR-generated DNA probe and three positive clones were isolated. Enzyme activities in E. coli clones were confirmed by bioassay and HPLC assay. Throughout the functional mapping, it was observed that the gene for isopenicillin N synthase is located at the 1.3-kb XhoI-BamHI fragment of insert of positive clones. Nucleotide sequencing at both ends of the XhoI-BamHI fragment revealed that IPNS of L. lactamgenus has the common amino acid sequences at amino- and carboxy-termini.

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Genetic Architecture of Transcription and Chromatin Regulation

  • Kim, Kwoneel;Bang, Hyoeun;Lee, Kibaick;Choi, Jung Kyoon
    • Genomics & Informatics
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    • v.13 no.2
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    • pp.40-44
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    • 2015
  • DNA microarray and next-generation sequencing provide data that can be used for the genetic analysis of multiple quantitative traits such as gene expression levels, transcription factor binding profiles, and epigenetic signatures. In particular, chromatin opening is tightly coupled with gene transcription. To understand how these two processes are genetically regulated and associated with each other, we examined the changes of chromatin accessibility and gene expression in response to genetic variation by means of quantitative trait loci mapping. Regulatory patterns commonly observed in yeast and human across different technical platforms and experimental designs suggest a higher genetic complexity of transcription regulation in contrast to a more robust genetic architecture of chromatin regulation.

Bacillus cellulyticus K-12 Crystalline Cellulose-Degrading Avicelase Gene and Expression in Eschterichia coli

  • Cheorl-Ho Kim;Woo
    • The Korean Journal of Food And Nutrition
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    • v.6 no.4
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    • pp.314-321
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    • 1993
  • We have cloned the Bacillus cellulyticus K-12 avicelase (Avi, E.C.3.2.1.4) gene (ace A) In E. coli. This was accompanied by using the vector PT7T3U 19 and Hind W -Hind m libraries of Bacillus cellulyticus K-12 chromosomal inserts created in 5.cofi. The Libraries were screened for the expression of avicelase by monitoring the immunoreaction of the anti-avicelase (immunoscreening). Positive clones (Ac-3, Ac-5, and Ac-7) contained the identical 3.5kb Hind III fragment as determined by restriction mapping and Southern hybridization, and expressed avicelase efficiently and constituvely using its own promoter in the heterologous host. From the immunoblotting analysis, a polypeptide which showed a CMCase activity with an Mr of 54000 was detected.

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Cloning and Characterization of Directly Amplified Antiviral Gene Interferon Alpha-2b (HulFN$\{alpha}$-2b) from Human Leukocytes Chromosomal DNA

  • Behravan, Javad;Ahmadpour, Hassan
    • Archives of Pharmacal Research
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    • v.27 no.7
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    • pp.776-780
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    • 2004
  • Interferons are cytokines that confer resistance to viral infection and inhibit cellular proliferation. The interferon alpha gene from human blood samples was amplified, cloned and expressed in E. coli (BL21). Leukocyte chromosomal DNA was used as a source of template DNA. Using specific primers, the gene for HulFN$\{alpha}$-2b was amplified and inserted into the E. coli vector, pET21b, by ligation of the HindIII and BamHI linkers of the vector and insert. The insert was further analyzed by PCR, DNA restriction mapping and sequencing, and expressed in a suitable E. coli strain. The production of this important cellular protein in the laboratory has significant applications in production of the recombinant pharmaceutical proteins.