Protein Tagging and Detection With Engineered Self - Assembling Fragments of Green Fluorescent Protein  

Oleh:

Cabantous, Stephanie ; Terwilliger, Thomas C. ; Waldo, Geoffrey S.

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Nature Biotechnology: The Science and Business of Biotechnology vol. 23 no. 1 (Jan. 2005), page 102-107.
Topik: PROTEIN; protein; fluoresecent

Ketersediaan

Perpustakaan Pusat (Semanggi) Nomor Panggil: NN9.2 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

Lihat Detail Induk

Isi artikel

Existing protein tagging and detection methods are powerful but have drawbacks. Split protein tags can perturb protein solubility or may not work in living cells. Green fluorescent protein (GFP) fusions can misfold or exhibit altered rocessing. Fluorogenic biarsenical environment and cell transfection or permeabilization. An ideal protein tag in vitro, would provide a sensitive analytical signal and would not require external chemical reagents or substrates. One way to accomplish this might be with a split GFP but the GFP fragments reported thus far are large and fold poorly require chemical ligation or fused folded and fluorescent GFP. We have engineered soluble, self associating fragments of GFP that can be used to tag and detect either soluble or insoluble proteins in living cells or cell lysates. The split GFP system is simple and does not change fusion protein solubility.

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