• Title/Summary/Keyword: Cytoplasmic hybrid

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Investigation of Cytoplasmic Male Sterility and Development of Maintainer and Restorer Lines in Rye (Secale cereale L.)

  • Heo, Hwa-Young;Hong, Byung-Hee;Seong, Rak-Chun;Park, Moon-Woong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.5
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    • pp.347-351
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    • 2000
  • Rye has been a major winter forage crop in Korea. Varietal improvement of rye has been practiced either by hybrid or population breeding systems. Hybrid breeding offers important advantages over population breeding since it is normally a cross-pollinated crop. The hybrid breeding in rye has been possible since cytoplasmically inherited forms of male sterility (CMS) and corresponding nuclear restorer genes were found. The objectives of this research were to develop the maintainer and restorer lines of Korean inbred lines and to estimate the effect of 'Pampa' type of CMS cytoplasm on yield and its related characteristics. For easy discrimination of male-sterile status of plants, anther scoring and the restore index system in which seed-setting and pollen quantity of viability were taken into account were established. High significant correlation between pollen quantity and pollen viability was found. For "Pampa" cytoplasm, four of 14 Korean inbred lines tested turned out to be a maintainer but no restorer was found. But for "235b" CMS cytoplasm, seven inbred lines acted as complete restorers. The Korean inbred rye lines acted mainly as maintainers in "Pampa" cytoplasm but acted mainly as restorer in "235b" cytoplasm. The 'Pampa' cytoplasm inducing male sterility reduced cohn length and plant height and increased the number of tiller, so forage yield and grain yield were enhanced. However, heading date was slightly delayed compared to the normal cytoplasm.elayed compared to the normal cytoplasm.

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Studies on the Pathogenicity of Cytoplasmic Polyhedrosis Virus and the Phenomena of Induction and Interference by Oral Inoculation of Foreign Virus to the Silkworm, Bombyx mori L. (가잠에 있어서 세포질다각체 바이러스의 병원성 및 이형 바이러스 접종에 의한 유발 간섭현상에 관한 연구)

  • 김근영;문재유;임종성;정태암
    • Journal of Sericultural and Entomological Science
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    • v.17 no.2
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    • pp.101-118
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    • 1975
  • The pathogenicity of hexagonal and tetragonal cytoplasmic polyhedron virus to one of present leading silkworm varieties, Jam 103${\times}$Jam 104, and its parents, Jam 103 and Jam 104, was investigated. The activation of occult virus by oral inoculation of foreign virus as well as the interference phenomena between the activated and inoculated cytoplasmic polyhedron viruses was observed and the results obtained are as follows. 1. The pathogenicity of cytoplasmic polyhedrosis viruses. 1) There was a high significant difference in the pathogenicity of hexagonal polyhedron virus between the hybrid and its parents showing the lowest infection rate in the hybrid (Jam 103${\times}$Jam 104), medium infection rate in the Japanese line (Jam 103) and the highest infection rate in the Chinese line (Jam 104). In the pathogenicityof tetragonal polyhedron virus, a significant difference was observed only between the hybrid (Jam 103${\times}$Jam 104) and the Chinese line (Jam 104) by showing a higher infection rate in the Chinese line than in the hybrid. 2) The pathogenicity of both hexagonal and tetragonal polyhedron viruses showed a high significant difference in different silkworm instars inoculated by showing a higher infection rate at the second instar than at the fourth instar. 3) The pathogenicity of both hexagonal and tetragonal polyhedron viruses was increased as the concentration of viruses inoculated increased. 2. The phenomena of induction and interference by oral inoculation of foreign virus. 1) The induction rate of cytoplasmic polyhedrosis virus was higher in the parents than in the hybrid. In the parents. a higher rate in the Chinese line than in the Japanese line was observed. 2) The effect of inoculation at different instars on the induction was studied and the induction rate was higher at the second instar inoculated than at the fourth instar inoculated. 3) The degree of activation of hexagonal polyhedron virus with inoculation of tetragonal polyhedron virus was very high when a lower concentration of virus was inoculated and it was very low when a higher concentration of virus was inoculated 4) The degree of activation of tetragonal polyhedron virus with inoculation of hexagonal polyhedron virus was very low when a lower concentration of virus was inoculated and it was not observed when a higher concentration of virus was inoculated. 5) The mixed infection rate with hexagonal and tetragonal polyhedron viruses was higher at the second instar inoculated than at the fourth instar inoculated. 6) It was observed that the secondary hexagonal pc]yhedron virus activated interfered with the primary tetragonal polyhedron virus inoculated when the inoculated concentration of the primary virus is low and the primary virus inoculated interfered with the secondary virus activated when the inoculated concentration of the primary virus is high.

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Functional and Physical Interaction between Human Lactate Dehydrogenase B and $Na^+/H^+$ Exchanger Isoform 1

  • Kim, Eun-Hee
    • Animal cells and systems
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    • v.13 no.3
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    • pp.283-288
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    • 2009
  • The ubiquitous plasma membrane $Na^+/H^+$ exchanger 1 (NHE1) is rapidly activated in response to various extracellular stimuli and maintains normal cytoplasmic pH. Yeast two-hybrid screening was used in order to identify proteins interacting with NHE1 using its cytoplasmic domain as a bait from HeLa cDNA library. One of the interacting cDNA clones was human Lactate dehydrogenase B (LDHB). In vitro translated LDHB was pulled down together with GST-NHE1.cd protein in the GST pull down assay, confirming the interaction in vitro. LDHB antibody immunoprecipitated endogenous LDHB together with NHE1 from H9c2 cells, validating cellular interaction between NHE1 and LDHB. Subsequent analysis revealed that the overexpression of LDHB increased intracellular PH, implying opening of the NHE1 transporter. Moreover, overexpression of LDHB activated caspase 3 and induced cell death, consistent with the expected phenotype of hyper-activation of NHE1. Collectively, our data indicate that LDHB modulates NHE1 activity via physical interaction.

Direct and functional interaction between dopamine D2 receptor and ALY

  • Yang, Jee-Hyeo;Kim, Hyun-Jin;Cheong, Da-Woon;Kim, Kyeong-Man
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.89.1-89.1
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    • 2003
  • The signaling pathway of dopamine D$_2$ receptor was studied using yeast two-hybrid system. The 3rd cytoplasmic loop of rat D$_2$ receptor was fond to interact with ALY. The interaction in the yeast was observed only with the 3rd cytoplasmic loop of D$_2$ receptor but not with that of D$_3$ or D$_4$ dopamine receptor. The interaction between two proteins was also confirmed by GST pull-down assay. Co-expression of D$_2$ receptor and ALY enhanced the expression of Lef-1 promoter in C6 cells and the promoter of D$_2$ dopamine receptor itself.

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Direct and functional interaction between dopamine D2 receptor and ALY

  • Yang, Ji-Hye;Cheong, Da-Woon;Seo, Hyung-Ju;Kim, Moon-Soo;Kim, Kyeong-Man
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.270.1-270.1
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    • 2002
  • The signaling pathway of D2 dopamine receptor was studied using yeaslt two-hybrid system.. The 3rd cytoplasmic loop of rat D2 dopamine receptor was used to screen the cDNA library of mouse brain. and ALY was found to interact with it. The interaction in the yeast was observed only with the 3rd cytoplasmic loop of D2 dopamine receptor but not with that of D3 or D4 dopamine receptor. The interaction between two proteins was also confirmed by GST pull-down assay. (omitted)

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Fermentation and Purification of LacZ-Fused Single Chain Insulin Precursor for($B^{30}$-Homoserine) Human Insulin

  • SeungYup Lee;Jeo
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.1 no.1
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    • pp.9-12
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    • 1996
  • In order to produce the single chain precursor of a novel human insulin analogue, (B30-Homoserine) insulin, the fermentative behaviors of Escherichia coli JM103 were studied, which harbors pKBA plasmid carrying a hybrid gene in which the gene for a single chain precursor was fused with lacZ gene under tac promoter. The maximal induction of gene expression was achieved when more than 0.05 mM of isopropyl-$\beta$-D-thiogalactopyranoside(IPTG) was supplemented to fermentation medium after 4 h cultivation of E. coli, and followed by longer than 2-h fermentation. The hybrid protein of the single chain insulin precursor was isolated from cytoplasmic inclusion bodies by dissolving in 8M urea solution, and purified through DEAE-Sephacel and Sephadex G-200 column chromatographies with a recovery of 35%. The finally purified hybrid protein showed a single band on sodium dodecyl sulfate-polyacrylamide gel.

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Screening of Transcriptional Regulator of the Draf Proto-oncogene Using the Yeast One-hybrid System

  • Park, So-Young;Park, Na-Hyun;Kwon, Eun-Jeong;Yoo, Mi-Ye
    • Journal of Life Science
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    • v.9 no.2
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    • pp.52-56
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    • 1999
  • The Raf, a cytoplasmic serine/thereonine protein kinase, acts as an important mediator of signals involving cell proliferation, differentiation and development. Multiple regulatory elements should participate in the expression of D-raf, Drosophila homolog of human c-raf-1. In order to search regulatory factors involved in the D-raf promoter activation, we accomplished the yeast one-hybrid screening using D-raf promoter region from bp-330 to -309 with respect to the transcription initiation site as bait. After screening, sixteen independent positive clones of ${\beta}$-galactosidase activties were identified and sequenced. Two clones having 94-98% identity with daughterless and one clone having 93% identity with escargot by Blast search among these clones were screened.

Isolation of the Gene for HIV-1 gp41 Interacting Protein (HIV gp41의 세포내 부분과 상호작용하는 단백질 유전자의 분리)

  • Kim, Eun-Mi;Kim, Jung-Woo
    • The Journal of Natural Sciences
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    • v.10 no.1
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    • pp.27-32
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    • 1998
  • To find the interacting protein with the cytoplasmic domain of HIV-1 gp41, the yeast two hybrid system was used for the expression cloning. Among the $1.4 \times 10^6 colonies, 20 colonies were selected as the final candidate for the interacting protein gene. The nucleotide sequencing revealed three kinds of protein, acidic ribosomal protein P0, beta tubulin, alpha catenin. These proteins interacted with the gp41 specifically in yeast system.

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Development of Gene-based Markers for the Allelic Selection of the Restorer-of-fertility Gene, Rfo, in Radish (Raphanus sativus)

  • Kim, Sunggil;Lim, Heerae;Cho, Kang-Hee;Park, Pue Hee;Park, Suhyung;Sung, Soon-Kee;Oh, Daegeun;Kim, Ki-Taek
    • Korean Journal of Breeding Science
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    • v.41 no.3
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    • pp.194-204
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    • 2009
  • Cytoplasmic male sterility (CMS) and fertility restoration have been utilized as valuable tools for $F_1$-hybrid seed production in many crops despite laborious breeding processes. Molecular markers for the selection of CMS-related genes help reduce the expenses and breeding times. A previously reported genomic region containing the Ppr-B gene, which is responsible for restoration of fertility and corresponds to the Rfo locus, was used to develop gene-based or so-called "functional" markers for allelic selection of the restorer-of-fertility gene (Rfo) in $F_1$-hybrid breeding of radish (Raphanus sativus L.) Polymorphic sequences among Rfo alleles of diverse breeding lines of radish were examined by sequencing the Ppr-B alleles. However, presence of Ppr-B homolog, designated as Ppr-D, interferes on specific PCR amplification of Ppr-B in certain breeding lines. The organization of Ppr-D, resolved by genome walking, revealed extended homology with Ppr-B even in the promoter region. Interestingly, PCR amplification of Ppr-D was repeatedly unsuccessful in certain breeding lines implying the lack of Ppr-D in these radishes. Ppr-B could only be successfully amplified for analysis through designing primers based on the sequences unique to Ppr-B that exclude interference from Ppr-D gene. Four variants of Rfo alleles were identified from 20 breeding lines. A combination of three molecular markers was developed in order to genotype the Rfo locus based on polymorphisms among four different variants. These markers will be useful in facilitating $F_1$-hybrid cultivar development in radish.

Development of Tobacco Mosaic Virus (TMV) resistant Flue-cured $\textrm{F}_1$ Hybrid, KF118

  • Chung, Yun-Hwa;Keum, Wan-Soo;Park, Sang-Ju;Jung, Suk-Hum;Kim, Yong-Am;Kang, Yue-Gyu;Chung, Youl-Young;Cho, Soo-Heon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.5
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    • pp.388-391
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    • 2003
  • Tobacco mosaic virus (TMV), TMV-common strain, causes severe damage at growth, yield and leaf quality on flue-cured tobacco in Korea. Controlling TMV outbreak with cultivating practices and chemicals are limited, thus, the best way would be developing TMV resistant variety. A new flue-cured hybrid, KF118, was developed from a cross of maternal parent MSTC175 and TC853 at the Agro-tech. Research Group, KT&G Central Research Institute. KF118 exhibited more vigorous growth characteristics in early stage than that of NC82, standard variety in Korea, while other characters were favorable to those of NC82, It is highly resistant to TMV and bacterial wilt(Ral-stonia solanacerum), and black shank(phytophthora parasitica var. nicotianae) is comparable to that of NC82. It should adapt well to the flue-cured production area, and can reduce premature flowering under unfavorable weather conditions. Yield of KF118 is 1 to 2% higher, and leaf quality is 4% higher than NC82. Fl hybrid, KF118, met acceptable standards for chemical and physical characteristics of cured leaf, and ranked high in good smoking taste by panel members in KT&G.