• Title/Summary/Keyword: Recombinant Protein Production

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Construction of Transfer Vector for Production of Baculovirus Occlusion Bodies that Contain Novel Recombinant Crystal Protein

  • Shim, Hee-Jin;Choi, Jae-Young;Roh, Jong-Yul;Li, Ming Shun;Je, Yeon Ho
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.118-119
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    • 2003
  • Baculovirus occlusion bodies have been recently engineered to incorporate foreign protein such as the Bacillus thuringiensis (Bt) CrylAc protein for improvement of insecticidal activity. In this study, polyhedrin, cylAc, egfp and crylCa genes were fused to produce occlusion bodies that contain novel recombinant crystal protein by homologous recombination between cylAc and crylCa genes in insect cells. (omitted)

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재조합 Escherichia coli 시스템을 이용한 재조합 말라리아 항원의 발현 최적화 연구

  • Hong, Seong-Hui;Park, Do-Yeong;Hwang, Yeong-Bo;Park, Hyeon;Hwang, Hyeon-A
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.711-714
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    • 2001
  • The production of the recombinant Plasmodium vivax merozoite surface protein (PvMSP) has been investigated in the recombinant E.coli system. Experimental optimization of the culture conditions, such as the effect of initial pH, and operating temperature has been tried on the growth of recombinant E.coli and on the overproduction of the target foreign protein.

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Recombinant Goat VEGF164 Increases Hair Growth by Painting Process on the Skin of Shaved Mouse

  • Bao, Wenlei;Yin, Jianxin;Liang, Yan;Guo, Zhixin;Wang, Yanfeng;Liu, Dongjun;Wang, Xiao;Wang, Zhigang
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.9
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    • pp.1355-1359
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    • 2014
  • To detect goat vascular endothelial growth factor (VEGF)-mediated regrowth of hair, full-length VEGF164 cDNA was cloned from Inner Mongolia cashmere goat (Capra hircus) into the pET-his prokaryotic expression vector, and the recombinant plasmid was transferred into E. coli BL21 cells. The expression of recombinant $6{\times}his-gVEGF164$ protein was induced by 0.5 mM isopropyl thio-${\beta}$-D-galactoside at $32^{\circ}C$. Recombinant goat VEGF164 (rgVEGF164) was purified and identified by western blot using monoclonal anti-his and anti-VEGF antibodies. The rgVEGF164 was smeared onto the dorsal area of a shaved mouse, and we noted that hair regrowth in this area was faster than in the control group. Thus, rgVEGF164 increases hair growth in mice.

Applications of Yeast Flocculation in Biotechnological Processes

  • Domingues, Lucilia;Vicente, Antonio A.;Lima, Nelson;Teixeira, Jose A.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.4
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    • pp.288-305
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    • 2000
  • A review on the main aspects associated with yeast flocculation and its application in biotechnological processes is presented. This subject is addressed following three main aspects-the basics of yeast flocculation, the development of "new" flocculating yeast strains and bioreactor development. In what concerns the basics of yeast flocculation, the state of the art on the most relevant aspects of mechanism, physiology and genetics of yeast flocculation is reported. The construction of flocculating yeast strains includes not only the recombinant constitutive flocculent brewer's yeast, but also recombinant flocculent yeast for lactose metabolisation and ethanol production. Furthermore, recent work on the heterologous $\beta$-galactosidase production using a recombinant flocculent Saccharomyces cerevisiae is considered. As bioreactors using flocculating yeast cells have particular properties, mainly associated with a high solid phase hold-up, a section dedicated to its operation is presented. Aspects such as bioreactor productivity and culture stability as well as bioreactor hydrodynamics and mass transfer properties of flocculating cell cultures are considered. Finally, the paper concludes describing some of the applications of high cell density flocculating bioreactors and discussing potential new uses of these systems.e systems.

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Design and Expression of Recombinant Antihypertensive Peptide Multimer Gene in Escherichia coli BL21

  • Rao, Shengqi;Su, Yujie;Li, Junhua;Xu, Zhenzhen;Yang, Yanjun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1620-1627
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    • 2009
  • The design and expression of an antihypertensive peptide multimer (AHPM), a common precursor of 11 kinds of antihypertensive peptides (AHPs) tandemly linked up according to the restriction sites of gastrointestinal proteases, were explored. The DNA fragment encoding the AHPM was chemically synthesized and cloned into expression vector pGEX-3X. After an optimum induction with IPTG, the recombinant AHPM fused with glutathione S-transferase (GST-AHPM) was expressed mostly as inclusion body in Escherichia coli BL21 and reached the maximal production, 35% of total intracellular protein. The inclusion body was washed, dissolved, and purified by cation-exchange chromatography under denaturing conditions, followed by refolding together with size-exclusion chromatography and gradual dialysis. The resulting yield of the soluble GSTAHPM (34 kDa) with a purity of 95% reached 399 mg/l culture. The release of high active fragments from the AHPM was confirmed by the simulated gastrointestinal digestion. The results suggest that the design strategy and production method of the AHPM will be useful to obtain a large quantity of recombinant AHPs at a low cost.

Transformation of Terpene Synthase from Polyporus brumalis in Pichia pastoris for Recombinant Enzyme Production

  • An, Ji-Eun;Lee, Su-Yeon;Ryu, Sun-Hwa;Kim, Myungkil
    • Journal of the Korean Wood Science and Technology
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    • v.46 no.4
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    • pp.415-422
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    • 2018
  • Terpenoids have a wide range of biological functions and have extensive applications in the pharmaceutical, cosmetic, and flavoring industry. The white-rot fungus, Polyporus brumalis, is able to synthesize terpenoids via terpene synthase, which catalyzes an important step that forms a large variety of sesquiterpene products from farnesyl pyrophosphate (FPP). To improve the production of sesquiterpenes, the terpene synthase gene was isolated from Polyporus brumalis and was heterologously transformed into a Pichia pastoris strain. The open reading frame of the isolated gene (approximately 1.2 kb) was inserted into Pichia pastoris to obtain a recombinant enzyme. Five transformants were obtained and the expression of terpene synthase was analyzed at the transcript level by reverse transcription PCR (polymerase chain reaction) and at the protein level by SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis). Expression of the terpene synthase gene product was elevated in the transformants and as expected the molecular weight of the protein was approximately 45 kDa. These recombinant enzymes have potential practical applications and future studies should focus on their functional characterization.

Medium Optimization for Phytase Production by Recombinant Escherichia coli Using Statistical Experimental Design

  • Choi, Won-Chan;Oh, Byng-Chul;Kim, Hyung-Kwoun;Lee, Eun-Sook
    • Journal of Microbiology and Biotechnology
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    • v.12 no.3
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    • pp.490-496
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    • 2002
  • The production of E. coli WC7 phytase from a recombinant E. coli strain was optimized using a statistical experimental design approach. Two-level complete factorial designs with seven variables were used for the media optimization. In the first optimization step, the influence of disodium succinate, yeast extract, $K_2HPO_4,\;NH_4H_2PO_4,\;MgSO_4$, NaCl, and trace elements on phytase production was evaluated. As a result, disodium succinate, yeast extract, $NH_4H_2PO_4$, NaCl, and the trace elements were found to have a positive influence on the phytase production, while $K_2HPO_4\;and\;MgSO_4$ had a negative influence. In the second step, the concentrations of disodium succinate and yeast extract were further optimized using central composite designs. The maximum phytase activity obtained was 234 U/ml using 15.9 g/1 disodium succinate, 20 g/1 yeast extract, 5 g/1 K_2HPO_4,\;10 g/1 NH_4H_2PO_4,\;1.5 g/1 MgSO_4$, 4 g/1 NaCl, and 1.5 m1/1 trace elements, which was about a 14-fold increase in comparison with that obtained using the basal medium.