Fourier transform infrared difference spectroscopy of rhodopsin mutants: light activation of rhodopsin causes hydrogen-bonding change in residue aspartic acid-83 during meta II formation.
Fourier transform infrared difference spectroscopy of rhodopsin mutants: light activation of rhodopsin causes hydrogen-bonding change in residue aspartic acid-83 during meta II formation.
复制标题
视紫红质突变体的傅里叶变换红外差光谱：视紫红质的光活化导致meta II形成过程中残基天冬氨酸83的氢键变化。
DOI：
10.1021/bi00090a001
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发表时间：
1993
期刊：
影响因子：
2.9
通讯作者：
Rothschild,KJ
中科院分区：
文献类型：
--
作者：
Rath,P;DeCaluwé,LL;Bovee-Geurts,PH;DeGrip,WJ;Rothschild,KJ
野生型和突变型牛视紫红质的制备与纯化。所有涉及视紫红质的操作均在弱红光下进行（Schott-Jena, RG 645）。按照描述进行了重组杆状病毒的定点诱变、克隆、生产和繁殖（Summers & Smith, 1987； Janssen等人，1988,1990,1991;decaluw<s:1>等人，1993）。在TNH-FH + 10%胎牛血清（FCS）、50 g/mL链霉素和50单位/mL青霉素的培养基中，将夜蛾IPLB-Sf9细胞系维持在27℃。在悬浮培养（100-1000）中实现了重组视蛋白（V-ops）的大规模生产
MATERIALS AND METHODSProduction and Purification of Wild-Type and Mutant Bovine Rhodopsin. All manipulations involving rhodopsin were performed in dim red light (Schott-Jena, RG 645). Sitedirected mutagenesis, cloning, production, and propagation of recombinant baculovirus were performed as described (Summers & Smith, 1987; Janssen et al., 1988, 1990, 1991; DeCaluwé et al., 1993). The Spodoptera frugiperda cell line IPLB-Sf9 was maintained at 27 C in TNH-FH medium plus 10% fetal calf serum(FCS), 50 g/mL streptomycin, and50 units/mL penicillin. Large-scale production of recombinant opsin (V-ops) was achieved in suspension culture (100-1000