摘要
采用室内实验技术,研究了直接从苏云金芽胞杆菌胞晶混合物提取原毒素的方法。胞晶混合物依次用0.5mol·L-1NaCl和无菌去离子水洗涤3次,洗涤后的沉淀物用0.05mol·L-1NaOH+0.1mmol·L-1PMSF溶液溶解10min,10000r·min-1离心15min去除不溶物。上清液以25%乙酸调至pH4.4,得原毒素蛋白沉淀,冷冻干燥。SDS-PAGE电泳扫描结果表明,原毒素分子量为130kDa,在提取过程中没有发生降解。该提取方法具有操作简便、成本低、效率高等优点。
The extraction method of protoxin of Bacillus thuringiensis directly from the mixtures of spores and crystals was studied. Crystals and spores of Bacillus thuringiensis were washed three times with 0.5 mol·L^-1 NaCl and three times with thrice distilled water. The precipitate was resuspended in 0.05 mol·L^-1 NaOH＋0.1 mmol·L^-1 PMSF for 10 min. The aqueous suspension was centrifuged at 10 000 r·min^-1 for 15 min （4℃） and insoluble materials were discarded. The pH of the supernatant liquor was adjusted to 4.4 by titration of 25% （v/v） acetic acid. The protoxin precipitate was obtained by centrifugation at 15 000 r ·min^-1 for 10 min and then lyophilised. The molecular weight of the protoxin was 130 kDa （SDS-PAGE） and no degradation of protoxin was observed during the purification. This method was noted for its advantages of simple operation, low cost and high efficiency.
出处
《农业环境科学学报》
CAS
CSCD
北大核心
2006年第2期535-536,共2页
Journal of Agro-Environment Science
基金
天津市自然科学基金(05YFJMJC05100)
关键词
苏云金芽胞杆菌
伴胞晶体
芽胞
原霉幕
Bacillus thuringiensos
parasporal crystal
spore
protoxin
