• Title/Summary/Keyword: Heterologous

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Inhibition of Yeast Growth by Broadly Cross-Reactive Antisera Elicited by Heterologous Mannan-Protein Conjugate

  • Machova, Eva;Korcova, Jana;Cizova, Alzbeta;Bystricky, Slavomir
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1177-1179
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    • 2015
  • A new approach to obtain broadly cross-reactive antisera against important yeast pathogens by intensive hyperimmunization with polysaccharide-protein conjugates is described here. Surface mannan of Candida albicans and capsular galactoglucoxylomannan of Cryptococcus laurentii were isolated and chemically linked to human serum albumin. Antisera elicited by a 7-week vigorous immunization of rabbits with the conjugates showed effective cross-reactive growth inhibition of different representatives of Candida spp. as well as Cryptococcus spp. IgG antibodies are evidenced as the effective component of the antisera.

Optimization of the Functional Expression of Coprinus cinereus Peroxidase in Pichia pastoris by Varying the Host and Promoter

  • Kim, Su-Jin;Lee, Jeong-Ah;Kim, Yong-Hwan;Song, Bong-Keun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.966-971
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    • 2009
  • Peroxidase from Coprinus cinereus (CiP) has attracted attention for its high specific activity and broad substrate spectrum compared with other peroxidases. In this study, the functional expression of this peroxidase was successfully achieved in the methylotrophic yeast Pichia pastoris. The expression level of CiP was increased by varying the microbial hosts and the expression promoters. Since a signal sequence, such as the alpha mating factor of Saccharomyces cerevisiae, was placed preceding the cDNA of the CiP coding gene, expressed recombinant CiP (rCiP) was secreted into the culture broth. The Mut Pichia pastoris host showed a 3-fold higher peroxidase activity, as well as 2-fold higher growth rate, compared with the $Mut^s $ Pichia pastoris host. Furthermore, the AOX1 promoter facilitated a 5-fold higher expression of rCiP than did the GAP promoter.

Heterologous Expression of Phanerochaete chrysoporium Glyoxal Oxidase and its Application for the Coupled Reaction with Manganese Peroxidase to Decolorize Malachite Green

  • Son, Yu-Lim;Kim, Hyoun-Young;Thiyagarajan, Saravanakumar;Xu, Jing Jing;Park, Seung-Moon
    • Mycobiology
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    • v.40 no.4
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    • pp.258-262
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    • 2012
  • cDNA of the glx1 gene encoding glyoxal oxidase (GLX) from Phanerochaete chrysosporium was isolated and expressed in Pichia pastoris. The recombinant GLX (rGLX) produces $H_2O_2$ over 7.0 nmol/min/mL using methyl glyoxal as a substrate. Use of rGLX as a generator of $H_2O_2$ improved the coupled reaction with recombinant manganese peroxidase resulting in decolorization of malachite green up to $150{\mu}M$ within 90 min.

Expression of heterologous genes using the slpA promoter and signal sequence in Lactobacilli

  • Gang, Seung-Ha;Bok, Jin-Deok;Jo, Gwang-Geun;Jo, Jae-Sun;Choe, Yun-Jae
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.202-205
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    • 2000
  • A gene coding $endo-{\beta}$,-1, 4 glucanase from Actinomyces sp. KNG40 and phytase from Hansenula polymorpha were cloned into Esherichia coli JM101 by using E. coli/Lactobacillus shuttle vector pNZ3004 and pNZ123. The plasmid p3PS(1-4) and p123(1-4) have slpA promoter and slpA signal sequence. So, I constructed expression vectors, p3PS(1-4)Endo, phy and p123(1-4)Endo, phy. These constructed vector was transformed in target host Lactobacillus gasseri and reutri. These transformed host expressed endoglucanase and phytase as extracellular fraction. In the enzyme activity of the same vector, host L, gasseri was higher activity than L. reuteri. This indicates that L. gasseri recongnize promoter and signal sequence very well.

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Effects of the Vitreoscilla Hemoglobin Gene on the Expression of the Ferritin Gene in Escherichia coli

  • Chung, Yun-Jo;Kim, Kyung-Suk;Jeon, Eun-Soon;Park, Kie-In;Park, Chung-Ung
    • BMB Reports
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    • v.31 no.5
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    • pp.503-507
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    • 1998
  • To investigate the effects of the Vitreoscilla hemoglobin (VHb) gene on the production of a heterologous protein, a comparative expression system for VHb and ferritin was constructed. First, the VHb gene was inserted into the downstream and upstream regions of the ferritin gene to construct pHF2 and pHF3, respectively. Next, the two plasmids pACHB1 and pVUTFH10, having the VHb gene and the ferritin gene respectively, were constructed in order to express the two genes in different plasmids by using a coplasmid expression system. It was observed that the cell growth was improved in all strains containing the VHb gene. Furthermore, in our coplasmid expression system, the presence of the VHb gene increased production of the ferritin by 1.8 times, as much as that in a strain not having the VHb gene.

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Cell cycle regulatory element in the promoter of the human thymidine kinase gene and its binding to factors

  • Kim, Yong-Kyu
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.10a
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    • pp.9-15
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    • 1995
  • When quiescent cells ate stimulated to enter the cell cycle, the thymidine kinase(TK) gene is transcriptionally activated at the border of Gl and 5. In this report we show that the human TK promoter contains multiple protein-binding sites. By site-directed mutagenesis, we identified a protein-binding site on the human TK promoter requited for conferring Gl-S-regulated transcription to a heterologous promoter and dissociated it functionally from an adjacent protein-binding domain containing an inverted CCAAT motif requited for high basal level expression. Substitution-mutation of this site results in constitutive expression of the neo reporter gene in serum-stimulated fibroblasts, as well as in cells arrested in mid-Gl by a temperature-sensitive mutation. The regulatory domains for the human TK promoter exhibit interesting symmetrical features, including a set of CCAAT motifs and sites similar to the novel Yi protein-binding site recently discovered in the mouse TK promoter. Thus, components of the hTK complex is important for hTK gene regulation.

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Super tree development by pyramiding heterologous functional genes

  • Noh, Eun-Woon
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.120-125
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    • 2005
  • Continuous degradation of forest in both quality and quantity threatens wood security in the future. Thus in the future, most wood and pulp will be expected to be produced from plantation forests. We attempt to produce superior trees suitable for such plantations with maximum productivity in limited land area. Tree productivity could be enhanced either by promoting growth and wood quality or by reducing loss caused by abiotic and biotic stresses. Genetic transformation techniques may offer ways to improve the productivity by enabling trees to tolerate the stresses or to covert limited resources into big biomass. With the availability of information on various functional genes and gene transfer techniques, it should be possible to develop such trees. In this presentation, our work to produce such trees at Korea Forest Research Institute is briefly introduced.

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Cell Separation of Vegetable Tissues by Protopectinase (Protopectinase를 이용한 식물조직의 단세포화)

  • 이승철;고보성;이대희;황용일
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.3
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    • pp.430-435
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    • 1997
  • Protopectinases are heterologous group of enzymes that degrades insoluble protopectin which consists of middle lamella between cells of plant tissues. Tissues of potato and carrot could be separated to single cells by addition of protopectinase isolated from Rhizopus sp. Color changes ofthe suspensions treated with protopectinase and mechanically macerated after 2 weeks at 4$^{\circ}C$, were investigated. Color change of the latter was very serious, however, that of the former was insignificant. Furthermore, after heat treatment at 121$^{\circ}C$ for 5min, the constituents of mechanically macerted carrot suspension were separated into two layers, but those of single celled carrot were not. Yields of carrot juices extracted from single celled suspensions and mechanical maceration were 93.6% and 56.0%, respectively. These results support that treatment of protopectinase can increase yield of juices extracted from plant, and manufacture high value-added products in food processing.

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Multilocus Genotyping to Study Population Structure in Three Buffalo Populations of India

  • Tantia, M.S.;Vijh, R.K.;Mishra, Bina;Kumar, S.T. Bharani;Arora, Reena
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.8
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    • pp.1071-1078
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    • 2006
  • Three buffalo populations viz. Bhadawari, Tarai and local buffaloes of Kerala were genotyped using 24 heterologous polymorphic microsatellite loci. A total of 140 alleles were observed with an average observed heterozygosity of 0.63. All the loci were neutral and 18 out of the 24 loci were in Hardy Weinberg Equilibrium. The $F_{IS}$ values (estimate of inbreeding) for 16 loci in all the three populations were negative. This indicated lack of population structure in the three populations. The effective number of immigrants was 5.88 per generation between the Tarai and Bhadawari populations which was quite high suggesting substantial gene flow. The genetic distances revealed closeness between the Tarai and Bhadawari populations which was expected from geographical contiguity. The FST values were not significantly different from zero showing no population differentiation. The Correspondence Analysis based on the allelic frequency data clustered the majority of the Tarai and Bhadawari individuals as an admixture.

Gene Expression and Secretion of Human Epidermal Growth Factor in a Methylotrophic Yeast Hansenula polymorpha (메나놀 자화 효모 Hansenula polymorpha를 이용한 재조합 인체 표피 성장인자 유전자의 발현 및 분비)

  • Oh, Yong-Ik;Sohn, Jung-Hoon;Choi, Eui-Sung;Kim, Hee-Chul;Rhee, Sang-Ki
    • Microbiology and Biotechnology Letters
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    • v.22 no.5
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    • pp.477-484
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    • 1994
  • Using a methylotrophic yeast Hansenula polymorpha, a heterologous gene expression and secretion system was developed for the production of hEGF(human Epidermal Growth Factor) which has been shown to promote epithelial cell proliferation and to inhibit gastric acid secretion. The hEGF gene was chemically synthesized according to the preferred codon usage in H. polymor- pha and expressed under the control of the strong and inducible methanol oxidase(MOX) promoter. The mating factor $\alpha$ pre-pro leader sequence of Saccharomyces cerevisiae was employed for hEGF to be secreted into the extracellular medium. This expression cassette was stably integrated into the host chromosomal DNA. Mature hEGF was efficiently expressed and secreted into the extracel- lular medium. About 24 mg/l of hEGF was detected in the cuture supernatant of a transformant with pA-EGF3 under the suboptimal culture conditions.

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