Improved metagenome screening efficiency by random insertion of T7 promoters

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dc.contributor.authorYu Jung Kim-
dc.contributor.authorHaseong Kim-
dc.contributor.authorSeo Hyeon Kim-
dc.contributor.authorEugene Rha-
dc.contributor.authorSu-Lim Choi-
dc.contributor.authorSoo Jin Yeom-
dc.contributor.authorH S Kim-
dc.contributor.authorSeung Goo Lee-
dc.date.accessioned2017-04-19T10:22:44Z-
dc.date.available2017-04-19T10:22:44Z-
dc.date.issued2016-
dc.identifier.issn01681656-
dc.identifier.uri10.1016/j.jbiotec.2016.05.018ko
dc.identifier.urihttps://oak.kribb.re.kr/handle/201005/13287-
dc.description.abstractMetagenomes constitute a major source for the identification of novel enzymes for industrial applications. However, current functional screening methods are hindered by the limited transcription efficiency of foreign metagenomic genes. To overcome this constraint, we introduced the 'Enforced Transcription' technique, which involves the random insertion of the bi-directional T7 promoter into a metagenomic fosmid library. Then the effect of enforced transcription was quantitatively assessed by screening for metagenomic lipolytic genes encoding enzymes whose catalytic activity forms halos on tributyrin agar plates. The metagenomic library containing the enforced transcription system yielded a significantly increased number of screening hits with lipolytic activity compared to the library without random T7 promoter insertions. Additional sequence analysis revealed that the hits from the enforced transcription library had greater genetic diversity than those from the original metagenome library. Enhancing heterologous expression using the T7 promoter should enable the identification of greater numbers of diverse novel biocatalysts from the metagenome than possible using conventional metagenome screening approaches.-
dc.publisherElsevier-
dc.titleImproved metagenome screening efficiency by random insertion of T7 promoters-
dc.title.alternativeImproved metagenome screening efficiency by random insertion of T7 promoters-
dc.typeArticle-
dc.citation.titleJournal of Biotechnology-
dc.citation.number0-
dc.citation.endPage53-
dc.citation.startPage47-
dc.citation.volume230-
dc.contributor.affiliatedAuthorHaseong Kim-
dc.contributor.affiliatedAuthorEugene Rha-
dc.contributor.affiliatedAuthorSeung Goo Lee-
dc.contributor.alternativeName김유정-
dc.contributor.alternativeName김하성-
dc.contributor.alternativeName김서현-
dc.contributor.alternativeName나유진-
dc.contributor.alternativeName최수림-
dc.contributor.alternativeName염수진-
dc.contributor.alternativeName김학성-
dc.contributor.alternativeName이승구-
dc.identifier.bibliographicCitationJournal of Biotechnology, vol. 230, pp. 47-53-
dc.identifier.doi10.1016/j.jbiotec.2016.05.018-
dc.subject.keywordEnforced-transcription-
dc.subject.keywordEsterase-
dc.subject.keywordGenetic diversity-
dc.subject.keywordHigh-efficiency screening-
dc.subject.keywordLipase-
dc.subject.keywordMetagenomic screening-
dc.subject.localEnforced-transcription-
dc.subject.localEsterase-
dc.subject.localGenetic diversity-
dc.subject.localHigh-efficiency screening-
dc.subject.localLipase-
dc.subject.localMetagenomic screening-
dc.description.journalClassY-
Appears in Collections:
Division of Biomaterials Research > Synthetic Biology and Bioengineering Research Center > 1. Journal Articles
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