摘要
粘虫颗粒体病毒经０．０２ｍｏｌ／ＬＮａＯＨ碱溶，先用ＳｅｐｈａｄｅｘＧ－２００凝胶过滤层析柱从病毒蛋白粗提物中分离增效因子，然后选用ＤＥＡＥ－ＳｅｐｈａｒｏｓｅＣＬ－６Ｂ离子交换层析柱进一步纯化增效因子，得到少量电泳纯的增效因子蛋白样品。纯化的增效因子具有典型的蛋白紫外吸收光谱，最大吸收波长为２７５．２ｎｍ。ＳＤＳ－ＰＡＧＥ电泳确定增效因子的分子量为１０８ｋＤ。用苏丹黑Ｂ脂蛋白染色法、过碘酸－Ｓｃｈｉｆ试剂糖蛋白染色法定性分析增效因子，非变性电泳凝胶上的增效因子带均为阴性反应。等电聚焦电泳检测增效因子的等电点为ｐＨ５．３左右。氨基酸组成分析证明，增效因子富含酸性氨基酸，占蛋白总量的２６．４２％。
The Hawaiian strain of Pseudaletia unipuncta GV was propagated in Pseudaletia separata larvae. The capsules of PuGV were dissolved in 0.02mol/L NaOH, the concentrated crude protein extract was firstly filtered through on a column of Sephadex G-200, the fraction mainly containing synergetic factor was further purified by applying onto a DEAE-Sepharose CL-6B column. The purified synergetic factor has typical ultraviolet absorptive spectrum of proteins and the wavelength, at which synergetic factor has maximum absorbance, is 275.2nm. Determined by SDS-PAGE, the molecular weight of synergetic factor is 108kD. Following native-PAGE, Sudan Black B staining method and Periodic acid-Schiff method were used to detect lipoprotein and protein-bound carbohydrates respectively, but the bands of synergetic factor had negative reactions. The isoelectric point of synergetic factor is pH5.3 analyzed by polyacrylamide gel isoelectric focusing. The results of amino acid analysis indicated that 26.42% of the purified synergetic factor consists of the acidic amino acids, aspartic and glutamic acids.
出处
《病毒学报》
CAS
CSCD
北大核心
1998年第4期352-358,共7页
Chinese Journal of Virology
基金
农业虫害鼠害综合治理研究国家重点实验室资助
微生物资源前期开发国家重点实验室部分资金资助
关键词
粘虫颗粒体病毒
增效因子
纯化
生化性质
Granulosis virus of Pseudaletia unipuncta, Synergetic factor, Purification, Biochemical characterization
