請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/69609
標題: Arabidopsis ENDO2: Its Catalytic Role and Requirement of N-Glycosylation for Function
作者: Ko, C.Y.
Lai, Y.L.
Liu, W.Y.
Lin, C.H.
Chen, Y.T.
Chen, L.F.O.
Lin, T.Y.
Shaw, J.F.
關鍵字: N-glycosylation
ENDO2
single-strand-preferring nuclease
mismatch
specific endonuclease
programmed cell death
programmed cell-death
strand-specific nucleases
penicillium-citrinum
substrate-specificity
p1 nuclease
dna
identification
endonuclease
purification
thaliana
期刊/報告no:: Journal of Agricultural and Food Chemistry, Volume 60, Issue 20, Page(s) 5169-5179.
摘要: The Arabidopsis thaliana At1g68290 gene encoding an endonuclease was isolated and designated ENDO2, which was cloned into a binary vector to overexpress ENDO2 with a C-terminal 6 x His-tag in A. thaliana. Our Arabidopsis transgenic lines harboring 35SP::ENDO2 produced stable active enzyme with high yield. The protein was affinity purified from transgenic plants, and its identity was confirmed by liquid chromatography-mass spectrometry and automatic Edman degradation. ENDO2 enzyme digests RNA, ssDNA, and dsDNA, with a substrate preference for ssDNA and RNA. The activity toward ssDNA (361.7 U/mg) is greater than its dsDNase activity (14.1 U/mg) at neutral pH. ENDO2 effectively cleaves mismatch regions in heteroduplex DNA containing single base pair mismatches or insertion/deletion bases and can be applied to high-throughput detection of single base mutation. Our data also validated that the removal of sugar groups from ENDO2 strongly affects its enzymatic stability and activity.
URI: http://hdl.handle.net/11455/69609
ISSN: 0021-8561
文章連結: http://dx.doi.org/10.1021/jf300945c
顯示於類別:期刊論文

文件中的檔案:
沒有與此文件相關的檔案。


在 DSpace 系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。