ID 111272
著者
Kogame, Toshiaki Max-Delbrück-Center for Molecular Medicine|The University of Tokushima
キーワード
Gateway cloning
MosSCI
C. elegans
transgenic
plasmid construct
資料タイプ
学術雑誌論文
抄録
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.
掲載誌名
The Journal of Medical Investigation
ISSN
13496867
13431420
cat書誌ID
AA11166929
AA12022913
出版者
Faculty of Medicine Tokushima University
62
3-4
開始ページ
161
終了ページ
166
並び順
161
発行日
2015-08
出版社版DOI
出版社版URL
フルテキストファイル
言語
eng
著者版フラグ
出版社版