• Title/Summary/Keyword: Gene order

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Use of the Non-electrophoretic Method to Detect Testis Specific Protein Gene for Sexing in Preimplantation Bovine Embryos

  • Huang, Jinming;You, Wei;Wu, Naike;Tan, Xiuwen
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.6
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    • pp.866-871
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    • 2007
  • Testis-specific protein (TSPY) is a Y-specific gene, with up to 200 copy numbers in bulls. In order to make bovine embryo sexing under farm condition more feasible, the possibility of using a non-electrophoretic method to detect the TSPY gene for sexing bovine early embryos was examined. Primers were designed to amplify a portion of the TSPY gene and a common gene as an internal control primer. PCR optimization was carried out using a DNA template from bovine whole blood. Furthermore, embryo samples were diagnosed by this method and the sexing results were contrasted with those of the Loop-Mediated Isothermal Amplification (LAMP) method. The results showed that TSPY was as reliable a sexing method as LAMP. Forty-three morula and blastocyst embryos collected from superovulated donor dairy cattle were sexed by this method, and twenty-one embryos judged to be female embryos were transferred non-surgically to recipients 6 to 8 days after natural estrus. Out of 21 recipients, 9 were pregnant (42.86%) and all delivered female calves. The results showed that the sex predicted by this protocol was 100% accurate. In conclusion, the TSPY gene was a good male specific marker and indicated that a non-electrophoretic method was feasible and accurate to detect the TSPY gene for sexing preimplantation bovine embryos.

Molecular cloning and sequence Analysis of the Gene for SecY from Streptomyces coelicolor (Muller) (Streptomyces coelicolor에서 secY 유전자의 클로닝과 염기서열 결정)

  • Kim, Sang-Suk;Hyun, Chang-Gu;Kim, Young-Min;Lee, Joo-Hun;Chung, In-Kwon;Kim, Dae-Myung;Suh, Joo-Won
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.678-686
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    • 1995
  • SecY is a central component of the protein export machinery that mediate the translocation of secretory proteins across the plasma membrane of Escherichia coli. In order to study the mechanism of protein secretion in Streptomyces, we have done cloning and sequencing of the Streptomyces coelicolor secY gene by using polymerase chain reaction method. The nucleotide sequence of the gene for SecY from S. coelicolor showed over 58% identity to that of M. luteus. The deduced amino acid sequences were highly homologous to those of other known SecY polypeptides, all having the potential to form 10 transmembrane segments, and especially second, fifth, and tenth segments were particularly conserved, sharing greater than 75% identity with W. lute s SecY. We propose that the conserved membrane-spanning segments actively participate in protein export. In B. subtilis and E. coli, the secY gene is a part of the spc operon, is preceded by the gene coding for ribosomal protein L15, and is likety coupled transcriptionally and translationally to the upstream L15 gene. In the other hand, secY gene of S. coelicolor and M. luteus have its own promoter region, are coupled translationally with adk gene and pr sented in adk operon.

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Granulosicoccaceae fam. nov., to Include Granulosicoccus antarcticus gen. nov., sp. nov., a Non-phototrophic, Obligately Aerobic Chemoheterotroph in the Order Chromatiales, Isolated from Antarctic Seawater

  • Lee, Ki-Young;Lee, Hong-Kum;Choi, Tae-Hwan;Kim, Kyung-Mi;Cho, Jang-Cheon
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1483-1490
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    • 2007
  • A Gram-negative, motile by tuft flagella, obligately aerobic chemoorganoheterotrophic, sphere-form bacterium, designated $IMCC3135^T$, was isolated from the Antarctic surface seawater of King George Island, West Antarctica. The strain was mesophilic, neutrophilic, and requiring NaCl for growth, but neither halophilic nor halotolerant. The 16S rRNA gene sequence analysis indicated that the strain was most closely related to genera of the order Chromatiales in the class Gammaproteobacteria. The most closely related genera showed less than 90% 16S rRNA gene sequence similarity and included Thioalkalispira (89.9%), Thioalkalivibrio (88.0%-89.5%), Ectothiorhodospira (87.9%-89.3%), Chromatium (88.3%-88.9%), and Lamprocystis (87.7%-88.9%), which represent three different families of the order Chromatiales. Phylogenetic analyses showed that this Antarctic strain represented a distinct phylogenetic lineage in the order Chromatiales and could not be assigned to any of the defined families in the order. Phenotypic characteristics, including primarily non-phototrophic, non-alkaliphilic, non-halophilic, and obligately aerobic chemoheterotrophic properties, differentiated the strain from other related genera. The very low sequence similarities (<90%) and distant relationships between the strain and members of the order suggested that the strain merited classification as a novel genus within a novel family in the order Chromatiales. On the basis of these taxonomic traits, a novel genus and species is proposed, Granulosicoccus antarcticus gen. nov., sp. nov., in a new family Granulosicoccaceae fam. nov. Strain $IMCC3135^T\;(=KCCM42676^T=NBRC\;102684^T)$ is the type strain of Granulosicoccus antarcticus.

Expression of Chitinase Gene in Solanum tuberosum L.

  • Park, Kyung-Hwa;Yang, Deok-Chun;Jeon, Jae-Heung;Kim, Hyun-Soon;Joung, Young-Hee;Hyouk Joung
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.85-90
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    • 1999
  • In order to protect fungal diseases, leaf disc explants of Solanum tuberosum cultivar, Belchip, was infected with an Agrobacterium MP90 strain containing chimeric gene construct, consisting of antibiotic resistance and chitinase gene driven by the CaMV 35S promoter, for transformation. Regenerated multiple shoots were selected on a medium containing kanamycin and carbenicillin after exposure to Agrobacterium. The presence and integration of the npt II and chitinase gene were confirmed by polymerase chain reaction(PCR). Northern blot analysis indicated that the genes coding for the enzyme could be expressed in potato plants. The chitinase activity of transgenic potato plants was higher than the control potato.

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Characteristics of Oncolytic Adenovirus Replication and Gene Expression in Hypoxic Condition

  • Kim, Hong-Sung
    • Biomedical Science Letters
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    • v.17 no.3
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    • pp.185-190
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    • 2011
  • Adenovirus type 5 (Ad5) vectors have been used for gene transfer to a wide variety of cell types in vivo and in vitro. The advantages of adenovirus vectors include the high titer of virus readily obtained in large scale preparations, their ability to transduce dividing and non dividing cells, and the high level of transgene expression. Since adenovirus vectors do not integrate in host cell DNA, there is a lack of insertional mutagenesis. However, many human tumor cells lack expression of the adenovirus 5 receptors and contain areas of hypoxia. In order to identify the pattern of replication and gene expression of oncolytic adenovirus in hypoxic condition, multiple different fiber modified Ads (Ad5F/S11, Ad5F/S35, Ad5F/K7, Ad5F/K21, and Ad5F/RGD) was compared. The replication of all fiber modified adenovirus was inhibited in hypoxic condition in HEK 293 cells, but gene expression has variety on different tumor cell lines and the level of coxackievirus and adenovirus receptor (CAR) expression. These data suggest that CAR expression pattern and hypoxic condition of tumor are considered for optimal oncolytic adenovirus application.

Cloning of the novel putative apoptosis-related gene of Spirometra erinacei (Order Pseudophyllidea)

  • Lee Soo-Ung;Huh Sun
    • Parasites, Hosts and Diseases
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    • v.44 no.3
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    • pp.233-237
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    • 2006
  • We postulated that apolysis was processed in accordance with apoptotic changes occurring in a cestode, Spirometra erinacei (Pseudophyllidea). We cloned the novel putative apoptosis-associated gene from S. erinacei via screening of a S. erinacei cDNA library with a ced-3 gene (activator of apoptosis) probe from Caenorhabditis elegans. We identified a 261-bp cDNA sequence, which encodes for an 86-amino acid protein. The cloned gene expression was observed in the neck and gravid proglottids via Northern blotting, using cloned cDNA inserts as probes, but the clone was not expressed in any of other tissues. We suggest that this gene may be involved in the apolysis of S. erinacei during normal tissue development and differentiation in cestode parasites.

Consensus Clustering for Time Course Gene Expression Microarray Data

  • Kim, Seo-Young;Bae, Jong-Sung
    • Communications for Statistical Applications and Methods
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    • v.12 no.2
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    • pp.335-348
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    • 2005
  • The rapid development of microarray technologies enabled the monitoring of expression levels of thousands of genes simultaneously. Recently, the time course gene expression data are often measured to study dynamic biological systems and gene regulatory networks. For the data, biologists are attempting to group genes based on the temporal pattern of their expression levels. We apply the consensus clustering algorithm to a time course gene expression data in order to infer statistically meaningful information from the measurements. We evaluate each of consensus clustering and existing clustering methods with various validation measures. In this paper, we consider hierarchical clustering and Diana of existing methods, and consensus clustering with hierarchical clustering, Diana and mixed hierachical and Diana methods and evaluate their performances on a real micro array data set and two simulated data sets.

Interaction of Heliothis armigera Nuclear Polyhedrosis Viral Capsid Protein with its Host Actin

  • Lu, Song-Ya;Qi, Yi-Peng;Ge, Guo-Qiong
    • BMB Reports
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    • v.35 no.6
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    • pp.562-567
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    • 2002
  • In order to find the cellular interaction factors of the Heliothis armigera nuclear polyhedrosis virus capsid protein VP39, a Heliothis armigera cell cDNA library was constructed. Then VP39 was used as bait. The host actin gene was isolated from the cDNA library with the yeast two-hybrid system. This demonstrated that VP39 could interact with its host actin in yeast. In order to corroborate this interaction in vivo, the vp39 gene was fused with the green fluorescent protein gene in plasmid pEGFP39. The fusion protein was expressed in the Hz-AM1 cells under the control of the Autographa californica multiple nucleopolyhedrovirus immediate early gene promoter. The host actin was labeled specifically by the red fluorescence substance, tetramethy rhodamine isothicyanete-phalloidin. Observation under a fluorescence microscopy showed that VP39, which was indicated by green fluorescence, began to appear in the cells 6 h after being transfected with pEGFP39. Red actin cables were also formed in the cytoplasm at the same time. Actin was aggregated in the nucleus 9 h after the transfection. The green and red fluorescence always appeared in the same location of the cells, which demonstrated that VP39 could combine with the host actin. Such a combination would result in the actin skeleton rearrangement.

Distribution and recombination of Wolbachia endosymbionts in Korean coleopteran insects

  • Jeong, Gilsang;Han, Taeman;Park, Haechul;Park, Soyeon;Noh, Pureum
    • Journal of Ecology and Environment
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    • v.43 no.4
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    • pp.421-426
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    • 2019
  • Background: Wolbachia are among the most prevalent endosymbiotic bacteria and induce reproductive anomalies in various invertebrate taxa. The bacterium has huge impacts on host reproductive biology, immunity, evolution, and molecular machinery. However, broad-scale surveys of Wolbachia infections at the order scale, including the order Coleoptera, are limited. In this study, we investigated the Wolbachia infection frequency in 201 Coleopteran insects collected in Korea. Results: A total of 26 species (12.8%) belonging to 11 families harbored Wolbachia. The phylogenetic trees of based on partial 16S rRNA gene sequences and partial Wolbachia surface protein (wsp) gene sequences were largely incongruent to that of their hosts. This result confirms that Wolbachia evolved independently from their hosts, Conclusion: Phylogenetic trees suggest that complex horizontal gene transfer and recombination events occurred within and between divergent Wolbachia subgroups.

Quantitative analysis of rutin content using silkworm genetic resources

  • Ju, Wan-Taek;Kim, Kee-Young;Sung, Gyoo-Byung;Kim, Yong-Soon
    • International Journal of Industrial Entomology and Biomaterials
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    • v.31 no.2
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    • pp.56-61
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    • 2015
  • Rutin is an important bioflavonoid that is consumed in the daily diet. This study compared the functional components of rutin from various silkworm species using a gene database with those of rutin produced by silkworms selectively bred through cross-combinations. We made comparisons between the geographical origin and species of silkworm using a gene database and discovered that rutin activity was ranked in the following order by species, Chinese (C5)> miscellaneous varieties (Jamsaeng 1 Ho) >Japanese (Jam 115) > European (E58) >Korean (Sun 3 ho). However, rutin levels with respect to various genetic traits (blood color, silk color, and egg color) were consistent. In order to study rutin changes that occurred during the cross breeding of the silkworm gene, we bred cross-combinations utilizing Jam 115 and the 4051 silkworms. In conclusion, in order to provide information about the constituents of functional materials contained in silkworm powder, it is imperative that silkworm cross breeding occurs so that the database of functional materials extracted from silkworms will expand.