• Title/Summary/Keyword: homologous

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Activation of the Caprine ${\beta}$-Lactoglobulin Gene Promoter by Lactogenic Hormones in Cultured Mammary HC11 Cells

  • Kim, Jae-Min;Yu, Myeong-Hui;Kim, Gyeong-Jin
    • Animal cells and systems
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    • v.1 no.4
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    • pp.603-608
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    • 1997
  • Analysis of the 5'-regulatory sequence of the caprine ${\beta}$-lactoglobulin (BLG) gene promoter revealed that two different types of activation were mediated by discrete regions, from -740 to -470 and from -205 to 109, in cultured mammary HC11 cells. Activation mediated by the proximal region was observed regardless of cell growth status. Distal activation, however, was observed only after confluent growth of the cells and was enhanced by the lactogenic hormones. This activation was accompanied by appearance of binding activity of proteins to these regions in the mammary HC11 cells. The binding motifs were broadly distributed over the upstream regulatory sequence. Comparison of the binding regions and mutation analysis suggest that a binding motif homologous to the ${\gamma}$-interferon responsive element (${\gamma}$-IRE) is responsible for transcriptional activation by hormonal induction in the mammary HC11 cells. The multiple ${\gamma}$-IRE homologous motifs seem to play a significant role in enhancing mammary cell-specific activation of the caprine BLG gene.

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Improvement of Glucoamylase Productivity of Saccharomyces diastaticus by Intergration of Glucoamylase Gene, STA, into Chromosomal DHA (Glucoamylase 유전자 STA의 염색체내 삽입에 의한 Saccharomyces diastaticus의 glucoamylase 생성능 향상)

  • 안종석;맹준호;강대욱;황인규;민태익
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.48-53
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    • 1993
  • For the purpose to improve the glucoamylase productivity of Saccharomyces diastaticus, we integrated STA 1 gene into chromosomal DNA of S. diastaticus using YIp vector. After construction of Ylp-STA by the subcloning of STAI (5.3 kb) into YIp5 vector, S. diastaticus GMT-II(a. ura3. STAJ) was transformed by Ylp-STA through homologous recombination at the chromosomal STAJ gene. So we obtained the tram formants that glucoamylase productivity was increased maximum six fold. These strains transformed by the multi-copy integration of Ylp-STA in chromosomal DNA were confirmed by Southern hybridization. And the integrated Ylp-STA was maintained stably during 30 mitotic divisions.

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Antigenicity of Partial Fragments of Recombinant Pasteurella multocida Toxin

  • Lee, Jeong-Min;Woo, Hee-Jong
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1756-1763
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    • 2010
  • Pasteurella multocida serogroup D strain, which produces P. multocida toxin (PMT), is a widespread and harmful pathogen of respiratory diseases such as pneumonia and progressive atrophic rhinitis (PAR) in swine. Vaccination has been considered the most desirable and effective approach for controlling the diseases caused by toxigenic P. multocida. To investigate the antigenicity and immunogenicity of partial fragments of recombinant PMT, recombinant proteins of the N-terminal (PMT-A), middle (PMT-B), C-terminal (PMT-C), and middle-C-terminal (PMT2.3) regions of PMT were successfully produced in an Escherichia coli expression system. The molecular masses of PMT-A, PMT-B, PMT-C, and PMT2.3 were ca. 53, 55, 35, and 84 kDa, respectively, purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. All the recombinant proteins except for PMT-A showed immune responses to antisera obtained from a swine showing symptoms of PAR. Moreover, high titers of PMT-specific antibodies were raised from mice immunized with each of the recombinant proteins; however, the immunoreactivities of the antibodies to authentic PMT and heat-inactivated whole bacteria were different, respectively. In the protection study, the highest protection against homologous challenge was shown in the case of PMT2.3; relatively poor protections occurred for the other PMT fragments.

Hed1 Promotes Meiotic Crossover Formation in Saccharomyces cerevisiae

  • Kong, Yoon-Ju;Joo, Jeong-Hwan;Kim, Keun Pil;Hong, Soogil
    • Journal of Microbiology and Biotechnology
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    • v.27 no.2
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    • pp.405-411
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    • 2017
  • Homologous recombination occurs between homologous chromosomes and is significantly involved in programmed double-strand break (DSB) repair. Activation of two recombinases, Rad51 and Dmc1, is essential for an interhomolog bias during meiosis. Rad51 participates in both mitotic and meiotic recombination, and its strand exchange activity is regulated by an inhibitory factor during meiosis. Thus, activities of Rad51 and Dmc1 are coordinated to promote homolog bias. It has been reported that Hed1, a meiosis-specific protein in budding yeast, regulates Rad51-dependent recombination activity. Here, we investigated the role of Hed1 in meiotic recombination by ectopic expression of the protein after pre-meiotic replication in Saccharomyces cerevisiae. DNA physical analysis revealed that the overexpression of Hed1 delays the DSB-to-joint molecule (JM) transition and promotes interhomolog JM formation. The study indicates a possible role of Hed1 in controlling the strand exchange activity of Rad51 and, eventually, meiotic crossover formation.

Identification and Characterization of Putative Integron-Like Elements of the Heavy-Metal-Hypertolerant Strains of Pseudomonas spp.

  • Ciok, Anna;Adamczuk, Marcin;Bartosik, Dariusz;Dziewit, Lukasz
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1983-1992
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    • 2016
  • Pseudomonas strains isolated from the heavily contaminated Lubin copper mine and Zelazny Most post-flotation waste reservoir in Poland were screened for the presence of integrons. This analysis revealed that two strains carried homologous DNA regions composed of a gene encoding a DNA_BRE_C domain-containing tyrosine recombinase (with no significant sequence similarity to other integrases of integrons) plus a three-component array of putative integron gene cassettes. The predicted gene cassettes encode three putative polypeptides with homology to (i) transmembrane proteins, (ii) GCN5 family acetyltransferases, and (iii) hypothetical proteins of unknown function (homologous proteins are encoded by the gene cassettes of several class 1 integrons). Comparative sequence analyses identified three structural variants of these novel integron-like elements within the sequenced bacterial genomes. Analysis of their distribution revealed that they are found exclusively in strains of the genus Pseudomonas.

Synthesis and in vitro cytotoxicity of a homologous series of 5-halosubstituted $1,3-Bis(\omega-cyanoalkyl)$uracil analogues

  • Kim, Jack-C.;Dong, Eun-Soo;Park, Jin-Il;Kim, Young-Hyeun;Choi, Soon-Kyu
    • Archives of Pharmacal Research
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    • v.19 no.1
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    • pp.62-65
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    • 1996
  • A homologous series of twenty, hitherto unreported, analogues of 5-halosubstituted $1, 3-Bis(\omega-cyanoalkyl)uracil$acyclic nucleosides were synthesized by the series of alkylation reactions of 5-halouracils with the corresponding chloroacetonitrile, chloropropionitrile, chlorobutyronitrile and 5-chlorovaleronitrile $(Cl-(C_ 2)_n-CN: n=l, 2, 3, 4)\; in\; anhydrous\; DMSO\; (or DMF)/K_2CO_3(or NaH)\; under\; 75^{\circ}C$ temperature. Antitumor activities for the synthesized compounds were determined against three cell lines (FM-3A cell, P-388 cell and U-938 cell lines). The compounds that exhibited moderate activity to significant activity, included la-b, 2a-b, 3a-c, and 4a, whose compounds were active against P-388, FM-3A and U-937 cell lines with the compounds la, lb, and 2a, showing significant antitumor activity (inhibitory concentrations $(IC_{50})$ ranged from 2.2 to $7.0\mug/ml$). Their strucrure-activity relationship did not show any activity differences in their effective chain length (methyl, ethyl, propyl, butyl) in 1, 3-bis(.omega.-cyanoalkyl) uracils.

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The Experimental Study on the Anti-allergic Effects of Leejungtang and Leejungtanggamibang (理中湯과 理中湯加味方의 抗알레르기 效果에 관한 實驗的 硏究)

  • Nam, Hae-Jeong;Chae, Byung-yoon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.13 no.1
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    • pp.60-77
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    • 2000
  • Experimental studies were done to research effects of Leejungtang and Leejungtanggamibang on the anti-allergic effects. We studied the vacular permeability response induced by histamine and serotonin, homologous PCA provoked by IgE-like antibody against EWA, contact dermatitis induced by picryl chloride and delayed type hypersensitivity response to SRBC. The results were obtained as follows: 1. In effects of the Leejungtang and Leejungtanggamibang on the vascular permeability responses to intradermal serotonin, the Leejungtang revealed insignificant effect and Leejungtanggamibang revealed significantly inhibitory effect. 2, In effects of the Leejungtang and Leejungtanggamibang on the vascular permeability responses to intradermal histamine, the Leejungtang revealed insignificant effect and Leejungtanggamibang revealed significantly inhibitory effect. 3. In the homologous PCA provoked by IgE-like antibody against white albumin, the Leejungtang revealed insignificant effect and Leejungtanggamibang revealed significantly inhibitory effect. 4. In the delayed type hypersensitivity responses to picryl chloride, the Leejungtang revealed insignificant effect and Leejungtanggamibang revealed significantly inhibitory effect. 5. In the delayed type hypersensitivity responses to sheep red blood cell the Leejungtang revealed insignificant effect and Leejungtanggamibang revealed significantly inhibitory effect.

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Superinfection exclusion of BVDV occurs not only at the level of structural protein -dispensable viral replication but also at the level of structural protein -required viral entry

  • Lee Y.-M.;Frolov I.;Rice C.M.
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2000.10a
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    • pp.66-77
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    • 2000
  • For a variety of viruses, the primary virus infection has been shown to prevent superinfection with a homologous secondary virus; however, the mechanism of exclusion has not been clearly understood. In this work, we demonstrated that BVDV -infected MDBK cells were protected from superinfection with a homologous superinfecting BVDV, one of the positive-sense RNA pestiviruses, but not with an unrelated rhabdovirus, such as vesicular stomatitis virus. Once superinfection exclusion was established by a primary infection with BVDV, the transfected infectious BVD viral RNA genome was shown to be competent for viral translation, but not viral replication. In addition, our results also demonstrated that upon superinfection, the. viral RNA genome of viral particles was not transferred into the cytoplasm of BVDV -infected cells. Using newly developed system involving rapid generation of the MDBK cells expressing BVD viral proteins, we subsequently found that expression of the viral structural proteins was dispensable for the block occurring at the level of viral RNA replication, but required for the exclusion at the level of viral entry step. Altogether, these findings provide evidence that the superinfection exclusion of BVDV occurs not only at the level of viral replication in which the viral replicase are involved, but also at the level of viral entry with which the viral structural proteins are associated, and that a cellular factor(s) play an essential role in this process.

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Optimum Conditions for Transformation of Synechocystis sp. PCC 6803

  • Zang, Xiaonan;Liu, Bin;Liu, Shunmei;Arunakumara, K.K.I.U.;Zhang, Xuecheng
    • Journal of Microbiology
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    • v.45 no.3
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    • pp.241-245
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    • 2007
  • This study was conducted to determine the optimal conditions for introduction of exogenous DNA into Synechocystis sp. PCC 6803. Of the three transformation techniques studied, electroporation, ultrasonic transformation and natural transformation, natural transformation showed the highest efficiency. Additionally, this study demonstrated that the higher plasmid concentration and longer homologous recombining fragments resulted in a greater number of transformants. For successful transformation, the lowest concentration of plasmid was $0.02\;{\mu}g/ml$, and the shortest homologous recombining fragment was 0.2 kb. Use of Synechocystis sp. PCC 6803 in the logarithmic growth phase resulted in two-fold higher transformation rate than that of the same organism when cells in the latent phase or the plateau phase were used for transformation. Pretreatment of the host strain, Synechocystis sp. PCC 6803, with EDTA (2 mM) for two days prior to transformation increased the transformation efficiency by 23%. Additionally, incubation of the cells and DNA for 5 h under light conditions increased the transformation efficiency by two orders of magnitude. Moreover, recovery treatment of the cells before they were plated onto antibiotic medium also increased the transformation efficiency.

Sequence Variations in the Non-Coding Sequence of CTX Phages in Vibrio cholerae

  • Kim, Eun Jin;Yu, Hyun Jin;Kim, Dong Wook
    • Journal of Microbiology and Biotechnology
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    • v.26 no.8
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    • pp.1473-1480
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    • 2016
  • This study focused on the variations in the non-coding sequences between ctxB and rstR of various CTX phages. The non-coding sequences of CTX-1 and CTX-cla are phage type-specific. The length of the non-coding region of CTX-1 and CTX-cla is 601 and 730 nucleotides, respectively. The non-coding sequence of CTX phage could be divided into three regions. There is a phage type-specific Variable region between two homologous Common regions (Common regions 1 and 2). The non-coding sequence of RS1 element is similar to CTX-1 except that Common region 1 is replaced by a short RS1-specific sequence. The non-coding sequences of CTX-2 and CTX-cla are homologous, indicating the non-coding sequence of CTX-2 is derived from CTX-cla. The non-coding region of CTX-O139 is similar to CTX-cla and CTX-2; however, it contains an extra phage type-specific sequence between Common region 2 and rstR. The variations in the non-coding sequences of CTX phages might be associated with the difference in the replication efficiency and the directionality in the integration into the V. cholerae chromosome.