Regulation of post-Golgi vesicle production in an in vitro system.
Regulation of post-Golgi vesicle production in an in vitro system.
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体外系统中高尔基体后囊泡产生的调节。
DOI：
10.1101/sqb.1995.060.01.021
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
DeLemos-Chiarandini,C
中科院分区：
文献类型：
--
作者：
Simon,JP;Ivanov,IE;Ren,M;Zeng,J;Shopsin,B;Hersh,D;Tempst,P;Erdjument-Bromage,H;Lui,M;DeLemos-Chiarandini,C
MDCK or HepG2 cells were grown, infected with VSV (Indiana Serotype) or influenza virus (strain A, PR8), pulse-labeled, and chased for 2 hours at 20~ as described previously (Gravotta et al. 1990). A Golgi fraction was prepared, essentially according to the method of Balch et al.(1984), and aliquots of this fraction were kept frozen in liquid nitrogen.Liver cytosolic protein fraction. A 1: 10 homogenate of the livers of starved Sprague-Dawley rats was prepared at 4~ in a buffer containing 0.2 M sucrose, 20 mM HEPES-KOH (pH 7.3), 1 mM DTF, 10 units/ml trasylol, and 1 mM PMSF. The proteins in a high-speed (2 hrs at 150,000 g in a Beckman TI60 rotor) supernatant were precipitated by addition of solid ammonium sulfate to 100% saturation, resuspended in homogenization buffer lacking sucrose, dialyzed against this buffer, and after clarification by centrifugation, concentrated to 60 mg protein per milliliter by ultrafiltration. The final cytosolic protein fraction was stored at-80~ in small aliquots.
