• Title/Summary/Keyword: gene localization

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Study of Electrophoretic Karyotypes of Fusarium Section Liseola (Fusarium 균의 section Liseola에 대한 핵형 연구)

  • 밍병례;안미선;최영길
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.192-196
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    • 1999
  • CHEF-PFGE(Contour-Clarnped Homogeneous Electric field- Pulsed Field Gel Electrophoresis) was used to identify electrophoretic karyotype for eight strains belonging to the Fzisoriuni section Liseolo. Chromosome numbers were nine to thirteen bands, ranging in size Cram 0.75 to 6.45 Mb. The total genome size was eslimated to range from 38.19 Mb to 43.12 Mb and numerous chromosome-length polymorphisms (CLPs) were observed. For the chromosome localizalion of the gene, 1GS sequence(2.6 Kb) of rDNA from F: moniliforme, chs-2 gene(2.8 Kb) and 4 - 3 gene(3.8 Kb) from Neuuospora cmssa were wed as probes.

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Cloning and Expression Analysis of a Novel Mouse Zinc Finger Protein Gene Znf313 Abundantly Expressed in Testis

  • Li, Na;Sun, Huaqin;Wu, Qiaqing;Tao, Dachang;Zhang, Sizhong;Ma, Yongxin
    • BMB Reports
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    • v.40 no.2
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    • pp.270-276
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    • 2007
  • We have cloned a novel mouse zinc finger protein gene Znf313 by rapid amplification of cDNA ends (RACE) according to the homologue of human ZNF313 gene. The cDNA is 2,163 base pairs (bp) in length and encodes a 229 amino acids (aa) protein with a $C_3HC_4$ ring finger domain and three $C_2H_2$ domains. 89% and 93% nucleotide (nt) and aa sequence identity is observed with its human homologue. Revealed by Northern blot and RT-PCR, full mRNA consists of 2.16 kb and widely expresses in tissues as a single transcript, most abundantly in heart, liver, kidney and testis. The expression of Znf313 in testis is detected in all development stages. Western blot analysis also reveals that Znf313 is expressed in the tissues. Immunohistochemical staining and subcellular localization demonstrate that Znf313 is expressed both in the cytoplasm and nucleus whereas predominantly localized in the nucleus. Present data suggests that Znf313 gene might play a fundamental role in gene transcription and regulation in organism and relates to spermatogenesis.

Tissue Specific Expression of Wound-Inducible RCaM-2 Promoter in Transgenic Tobacco Plants (상처에 의해서 유도되는 벼 calmodulin promoter의 transgenic 담배에서조직 특이적 발현)

  • Choi Young Ju
    • Journal of Life Science
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    • v.15 no.2 s.69
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    • pp.176-181
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    • 2005
  • To study calmodulin (CaM) gene expression and its regulation, rice CaM promoter (RCaM-2) was isolated and fused to $\beta-glucuronidase$ (GUS), reporter gene. X-Glue staining patterns revealed that GUS localization is high in meristemic tissues such as the stem apex, stolen tip, and vascular regions. GUS staining in the transverse sections of stem and petiole was restricted to the inside of the vascular system, and cortex and epidermis located outside of the vascular system usually did not show GUS staining even a plant that expressed strong activity. GUS activity was found to be tissue specific expressed and exhibited a dramatic transient increase in response to wounding. These results suggest that the 5'-flanking region of RCaM gene regulates wound-inducible expression.

Subcellular Localization of Diacylglycerol-responsive Protein Kinase C Isoforms in HeLa Cells

  • Kazi, Julhash U.;Kim, Cho-Rong;Soh, Jae-Won
    • Bulletin of the Korean Chemical Society
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    • v.30 no.9
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    • pp.1981-1984
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    • 2009
  • Subcellular localization of protein kinase often plays an important role in determining its activity and specificity. Protein kinase C (PKC), a family of multi-gene protein kinases has long been known to be translocated to the particular cellular compartments in response to DAG or its analog phorbol esters. We used C-terminal green fluorescent protein (GFP) fusion proteins of PKC isoforms to visualize the subcellular distribution of individual PKC isoforms. Intracellular localization of PKC-GFP proteins was monitored by fluorescence microscopy after transient transfection of PKC-GFP expression vectors in the HeLa cells. In unstimulated HeLa cells, all PKC isoforms were found to be distributed throughout the cytoplasm with a few exceptions. PKC$\theta$ was mostly localized to the Golgi, and PKC$\gamma$, PKC$\delta$ and PKC$\eta$ showed cytoplasmic distribution with Golgi localization. DAG analog TPA induced translocation of PKC-GFP to the plasma membrane. PKC$\alpha$, PKC$\eta$ and PKC$\theta$ were also localized to the Golgi in response to TPA. Only PKC$\delta$ was found to be associated with the nuclear membrane after transient TPA treatment. These results suggest that specific PKC isoforms are translocated to different intracellular sites and exhibit distinct biological effects.

Expression and localization of the spermatogenesis-related gene, Znf230, in mouse testis and spermatozoa during postnatal development

  • Song, Hongxia;Su, Dan;Lu, Pan;Yang, Jiyun;Zhang, Wei;Yang, Yuan;Liu, Yunqiang;Zhang, Sizhong
    • BMB Reports
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    • v.41 no.9
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    • pp.664-669
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    • 2008
  • Znf230, the mouse homologue of the human spermatogenesis-related gene, ZNF230, has been cloned by rapid amplification of cDNA ends (RACE). This gene is expressed predominantly in testis, but its expression in different testicular cells and spermatogenic stages has not been previously analyzed in detail. In the present study, the cellular localization of the Znf230 protein in mouse testis and epididymal spermatozoa was determined by RT-PCR, immunoblotting, immunohistochemistry and immunofluorescence. It is primarily expressed in the nuclei of spermatogonia and subsequently in the acrosome system and the entire tail of developing spermatids and spermatozoa. The results indicate that Znf230 may play an important role in mouse spermatogenesis, including spermatogenic cell proliferation and sperm maturation, as well as motility and fertilization.

Actin Cytoskeleton and Golgi Involvement in Barley stripe mosaic virus Movement and Cell Wall Localization of Triple Gene Block Proteins

  • Lim, Hyoun-Sub;Lee, Mi Yeon;Moon, Jae Sun;Moon, Jung-Kyung;Yu, Yong-Man;Cho, In Sook;Bae, Hanhong;DeBoer, Matt;Ju, Hojong;Hammond, John;Jackson, Andrew O.
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.17-30
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    • 2013
  • Barley stripe mosaic virus (BSMV) induces massive actin filament thickening at the infection front of infected Nicotiana benthamiana leaves. To determine the mechanisms leading to actin remodeling, fluorescent protein fusions of the BSMV triple gene block (TGB) proteins were coexpressed in cells with the actin marker DsRed: Talin. TGB ectopic expression experiments revealed that TGB3 is a major elicitor of filament thickening, that TGB2 resulted in formation of intermediate DsRed:Talin filaments, and that TGB1 alone had no obvious effects on actin filament structure. Latrunculin B (LatB) treat-ments retarded BSMV cell-to-cell movement, disrupted actin filament organization, and dramatically decreased the proportion of paired TGB3 foci appearing at the cell wall (CW). BSMV infection of transgenic plants tagged with GFP-KDEL exhibited membrane proliferation and vesicle formation that were especially evident around the nucleus. Similar membrane proliferation occurred in plants expressing TGB2 and/or TGB3, and DsRed: Talin fluorescence in these plants colocalized with the ER vesicles. TGB3 also associated with the Golgi apparatus and overlapped with cortical vesicles appearing at the cell periphery. Brefeldin A treatments disrupted Golgi and also altered vesicles at the CW, but failed to interfere with TGB CW localization. Our results indicate that actin cytoskeleton interactions are important in BSMV cell-to-cell movement and for CW localization of TGB3.

Immunofluorescence Localization of Schizosaccharomjyces pombe $cdc103^{+}$ Gene Product

  • Kim, Hyong-Bai
    • Journal of Microbiology
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    • v.34 no.3
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    • pp.248-254
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    • 1996
  • $cdc103^+$ gene in Schizosaccharomyces pombe which is similar to the CDC3 gene in Saccharomyces cerevisiae was cloned and sequenced. Comparison of the predicted amino acid sequences of $cdc103^+$ and CDC3 revealed that they share significant similarity (43% identity and 56% identity or similarity) to each other. The gene product of CDC3 in S. cerevisiae is known to be a highly ordered ring of filaments that lies just inside the cytoplasmic membrane in the region of the mother-bud neck. In order to characterize the gene product of $cdc103^+$ in Schizosaccharomyces pombe, fusion proteins were used to generate the polyclonal antibodies specific for the gene product (cdc103p). In immunofluorescence experiments, these antibodies decorate the region of the septum formation as a double ring structure late in the cell division cycle.

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The Optimization for Functional Expression of Arabidopsis Thaliana AtPIP2-1 in Xenopus laevis Oocyte (Xenopus oocyte에서 애기장대 AtPIP2-1 활성측정을 위한 발현 최적화 조건 규명)

  • Kim, Hyun-Mi;Hwang, Hyun-Sik;Lee, Suk-Chan;Jo, Su-Hyun;Kim, Beom-Gi
    • Journal of Applied Biological Chemistry
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    • v.53 no.4
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    • pp.189-194
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    • 2010
  • We confirmed the hypo-osmotic shock strengths and duration, different type of vectors, and subcelluar localization to identify the optimum analysis condition of plant aquaporin activity in Xenopus ooctye using Arabidopsis thaliana AtPIP2-1 gene. Six minutes and 1/5ND buffer hypoosmotic shock treatment was the best condition to show the maximum swelling of Xenopus oocytes where AtPIP2-1 was expressed using pcDNA3.1 vector. AtPIP2-1 protein was expressed more efficiently in pGEMHE vector which has 5' and 3' UTR (untranslation region) of Xenopus ${\beta}$-GLOBIN gene in multiple cloning site than in pcDNA3.1 vector. Also green fluorescence of GFP fused to AtPIP2-1 was detected onto oocyte plasmamembrane where is the proper subcellular localization target of AtPIP2-1.

Localization of MAK18 gene on chromosome VIII of saccharomyces cerevisiae (Saccharomyces cerevisiae의 염색체 VIII상의 MAK 18 유전자 국소화)

  • 윤순찬;이현숙;이창원
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.318-323
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    • 1988
  • MAK18 gene of Saccharomyces cerevisiae, needed for M1 replication, was mapped within 2cM of PET3 on chromosome VIII. From 38kb clone pRE66 carrying SPO11 and PET3, we have localized MAK18 gene whose insert is 2.8kb. MAK18 gene is Iocalized on about 9kb distance from PET3 and about 18kb distance from SPO11 on chromosome VIII.

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Localization of a Human-Specific Retroposon (SINE-R.C2) to Chromosome 6p21.31 by Radiation Hybrid Mapping

  • Kim, Heui-Soo;Timothy J. Crow
    • Journal of Life Science
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    • v.10 no.2
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    • pp.12-13
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    • 2000
  • A human-specific retroposon SINE-R.C2 has been derived from a human endogenous retrovirus HER V-K 10. It is absent in the genome of nonhuman primates and present within the third intron of the human C2 gene that is located in the class III region of the major histocompatibility complex. In the present study, we determined the regional location of the human C2 gene. The analysis of the Genebridge 4 radiation hybrid mapping panel using PCR amplification located the C2 gene between D6S1422 (10.1 cR) and CHLC.GATA4A03 (21.3) with a lod score of>3.0. This allowed us to localize C2 gene on the human chromosome 6 band p21.31.

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