Parry M E, Stow N D, Marsden H S
Medical Research Council Virology Unit, Institute of Virology, Glasgow, U.K.
J Gen Virol. 1993 Apr;74 ( Pt 4):607-12. doi: 10.1099/0022-1317-74-4-607.
A rapid and simple two-step scheme for the purification of herpes simplex virus type 1 UL8 protein from insect cells infected with a recombinant baculovirus has been developed. The scheme involves DEAE-Sepharose and phenyl-Sepharose chromatography and yields approximately 1.5 mg of protein from 2.4 x 10(8) infected cells. The protein remains intact during purification as judged by its reactivity with amino and carboxy termini-specific antisera. Gel filtration chromatography showed that the protein exists as a monomer in solution. No binding of the protein to ssDNA or dsDNA or to a DNA/RNA hybrid could be demonstrated using a gel mobility shift assay.
已开发出一种快速简便的两步法，用于从感染重组杆状病毒的昆虫细胞中纯化单纯疱疹病毒1型UL8蛋白。该方法包括DEAE-琼脂糖和苯基-琼脂糖层析，从2.4×10⁸个感染细胞中可获得约1.5mg蛋白质。根据其与氨基和羧基末端特异性抗血清的反应性判断，该蛋白在纯化过程中保持完整。凝胶过滤层析表明该蛋白在溶液中以单体形式存在。使用凝胶迁移率变动分析未证明该蛋白与单链DNA、双链DNA或DNA/RNA杂交体有结合。
