4-fragment gateway cloning format for MosSCI-compatible vectors integrating promoterome and 3'UTRome libraries of Caenorhabditis elegans

Item Type:	Article
Title:	4-fragment gateway cloning format for MosSCI-compatible vectors integrating promoterome and 3'UTRome libraries of Caenorhabditis elegans
Creators Name:	Kogame, T.
Abstract:	The technique of Mos1-mediated Single Copy Insertion (MosSCI) now has become the essential technique which facilitates transgenic experiments for Caenohabditis elegans (C. elegans). Gateway system which is adopted to MosSCI-compatible vectors offers an advantage of simultaneous cloning with entry vectors cloned in the Gateway system format. On the other hand, the format for MosSCI-compatible vectors restricts flexibility in designing the vectors to only 3-fragment integration. Thus, construct of complex transgene such as the expression vector for translational gene fusion is tedious work even with Gateway system. We have developed the new recombination format called LeGaSCI (Library-enhanced Gateway for MosSCI) to expand the conventional 3-fragment to 4-fragment format which still retains the capacity to accept Promoterome and 3'UTRome libraries of C. elegans. In the new recombination format, 2 different Gateway format were combined. Cloning reaction for the tissue-specific expression vector of GFP-tagged protein with 3'UTR successfully occurred without any expected insertion, deletion or frame-shift mutation. Moreover, The MosSCI transgenic line was successfully generated with the construct. Collectively, we established the new Gateway system format which allows us to assemble 4-fragment insertion with the widest variety of entry clone vectors from C. elegans libraries.
Keywords:	Gateway Cloning, MosSCI, C. elegans, Transgenic, Plasmid Construct, Animals, Caenohabditis elegans
Source:	Journal of Medical Investigation
ISSN:	1343-1420
Publisher:	University of Tokushima
Volume:	62
Number:	3.4
Page Range:	161-166
Date:	18 September 2015
Official Publication:	https://doi.org/10.2152/jmi.62.161
PubMed:	View item in PubMed

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