Rapid selection of multiple gene integrant for the production of recombinant hirudin in Hansenula polymorpha

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dc.contributor.authorHwa Young Kim-
dc.contributor.authorJung Hoon Sohn-
dc.contributor.authorChul Ho Kim-
dc.contributor.authorK Jagannadha Rao-
dc.contributor.authorEui Sung Choi-
dc.contributor.authorMyung Kuk Kim-
dc.contributor.authorSang Ki Rhee-
dc.date.accessioned2017-04-19T08:56:56Z-
dc.date.available2017-04-19T08:56:56Z-
dc.date.issued2000-
dc.identifier.issn12268372-
dc.identifier.urihttps://oak.kribb.re.kr/handle/201005/5041-
dc.description.abstractFor the rapid selection of higher recombinant hirudin producing strain in a methylotrophic yeast Hansenula polymorpha, a multiple gene integration and dose-dependent selection vector, based on a telomere-associated ARS and a bacterial aminoglycoside 3-phosphotransferase (aph) gene, was adopted. Two hirudin expression cassettes (HV1 and HV2) were constructed using the MOX promoter of H. polymorpha and the mating factor α secretion signal of S. cerevisiae. Multiple integrants of a transforming vector containing hirudin expression cassettes were easily selected by using an antibiotic, G418. Hirudin expression level and integrated plasmid copy number of the tested transformants increased with increasing the concentration of G418 used for selection. The expression level of HV1 was consistently higher than that of HV2 under the similar conditions, suggesting that the gene context might be quite important for the high-level gene expression in H. polymorpha. The highest hirudin producing strain selected in this study produced over 96 mg/L of biologically active hirudin in a 500-mL flask and 165 mg/L in a 5-L fermentor.-
dc.publisherSouth Korea-
dc.titleRapid selection of multiple gene integrant for the production of recombinant hirudin in Hansenula polymorpha-
dc.title.alternativeRapid selection of multiple gene integrant for the production of recombinant hirudin in Hansenula polymorpha-
dc.typeArticle-
dc.citation.titleBiotechnology and Bioprocess Engineering-
dc.citation.number1-
dc.citation.endPage6-
dc.citation.startPage1-
dc.citation.volume5-
dc.contributor.affiliatedAuthorJung Hoon Sohn-
dc.contributor.affiliatedAuthorChul Ho Kim-
dc.contributor.affiliatedAuthorEui Sung Choi-
dc.contributor.alternativeName김화영-
dc.contributor.alternativeName손정훈-
dc.contributor.alternativeName김철호-
dc.contributor.alternativeNameRao-
dc.contributor.alternativeName최의성-
dc.contributor.alternativeName김명국-
dc.contributor.alternativeName이상기-
dc.identifier.bibliographicCitationBiotechnology and Bioprocess Engineering, vol. 5, no. 1, pp. 1-6-
dc.identifier.doi10.1007/BF02932344-
dc.subject.keywordhirudin-
dc.subject.keywordHansenula polymorpha-
dc.subject.keywordG418 resistance-
dc.subject.keywordmulticopy integration-
dc.subject.keywordMOX promoter-
dc.subject.localhirudin-
dc.subject.localHirudin-
dc.subject.localHansenula polymorpha-
dc.subject.localG418 resistance-
dc.subject.localmulticopy integration-
dc.subject.localmulti-copy integration-
dc.subject.localMOX promoter-
dc.description.journalClassY-
Appears in Collections:
Division of Biomaterials Research > Synthetic Biology and Bioengineering Research Center > 1. Journal Articles
Jeonbuk Branch Institute > Microbial Biotechnology Research Center > 1. Journal Articles
Division of Bio Technology Innovation > BioProcess Engineering Center > 1. Journal Articles
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