Temperature Sensitive Control of Protein Activity by Conditionally Splicing Inteins  

Oleh:

Zeidler, Martin P. ; Tan, Change ; Bellaiche, Yohanns ; Cherry, Sara ; Hader, Sabine ; Gayko, Urte ; Perrimon, Norbert

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Nature Biotechnology: The Science and Business of Biotechnology vol. 22 no. 7 (Jul. 2004), page 871-876.
Topik: TEMPERATURE CONTROL; temperature sensitive control. protein activity; conditionally splicing inteins

Ketersediaan

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

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Isi artikel

Conditional or temperature sensitive (TS) alleles represent useful tools with which to investigate gene function. Indeed, much of our understanding of yeast has relied on temperature sensitive mutations which when available also provide important insights into other model systems. However the rarity of temperature sensitive alleles and difficulty in identifying them has limited their use. here we describe a system to generate temperature sensitive alleles based on conditionally active inteins. We have identified temperature sensitive splicing variants of the yeast saccharomyces cerevisiae vacuolar ATPase subunit (VMA) intein inserted within Gal4 and transferred these into Gal80. We show that Gal80-intein is able to efficiently provide temporal regulation of the Gal4 / upstream activation sequence (UAS) system in a temperatue dependent manner in drosophila melanogaster. Given the minimal host requirements necessary for temperature sensitive intein splicing, this technique has the potential to allow the generation and use of conditionally active inteins in multiple host proteins and model systems, thereby widening the use of temperature sensitive alleles for functional protein analysis.

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