Abstract:
:Insertion and deletion analyses of a protein have been less common than point mutation analyses, partly due to the lack in effective methods. This is the case with the green fluorescent protein (GFP), which is so widely applied in molecular biology and other fields. In this paper we first introduce a systematic approach for generating insertion/deletion mutants of GFP. A new technology of Y-ligation-based block shuffling (YLBS) was successfully applied to produce size-altered GFPs, providing insertion-containing GFPs of fluorescence, though no deletion type of fluorescence was obtained so far as examined. The analysis of these proteins suggested that size alteration (deletion/insertion) is acceptable so far as some type of rearrangement in a local structure can accommodate it. This paper demonstrates that YLBS can generate insertion and deletion mutant libraries systematically, which are beneficial in the study of structure-function relationship.
journal_name
FEBS Lettjournal_title
FEBS lettersauthors
Kitamura K,Yoshida C,Nishigaki Kdoi
10.1016/s0014-5793(03)01308-5keywords:
subject
Has Abstractpub_date
2003-12-18 00:00:00pages
483-8issue
3eissn
0014-5793issn
1873-3468pii
S0014579303013085journal_volume
555pub_type
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