상세 정보

underline
Metadata Downloads : dc(xml) or Excel
Cited 0 time in scopus ci

Title 

Improved metagenome screening efficiency by random insertion of T7 promoters

Authors 

Yu Jung KimHaseong KimSeo Hyeon KimEugene RhaSu-Lim ChoiSoo Jin YeomH S KimSeung Goo Lee

Publisher 

Elsevier

Issue Date 

2016

Citation 

Journal of Biotechnology, vol. 230, no. 0, pp. 47-53

Keywords 

Enforced-transcriptionEsteraseGenetic diversityHigh-efficiency screeningLipaseMetagenomic screening

Abstract 

Metagenomes 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.

ISSN 

0168-1656

Link 

http://dx.doi.org/10.1016/j.jbiotec.2016.05.018

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


There are no files associated with this item.
qrcode

FusionCharts.
DSpace Software Coptright(c) 2010 MIT and Hewleft-Packard  /  KRIBB-REPOSITORY ( Email:jakim@kribb.re.kr)