酒石酸钾-甘油密度梯度离心法纯化对虾肝胰腺细小病毒的方法学研究

METHODOLOGICAL STUDY OF THE PURIFICATION OF THE HEPATOPANCREATIC PARVOVIRUS OF PENAEID SHRIMP BY A POTASSIUM TARTRATE-GLYCEROL DENSITY GRADIENT CENTRIFUGATION

  • 摘要: 阐述一种从对虾肝胰腺组织中纯化肝腺细小病毒的方法。将对虾肝胰腺组织样品 研磨并进行适当脱脂处理后, 进行一系列的浓缩和过35%( w / w ) 蔗糖垫离心, 然后通过一个不连续 的酒石酸钾- 甘油密度梯度离心纯化。结果证实, 该方法对样品中的病毒颗粒的分离和纯化效果均 较理想, 可以满足有关研究工作的需要。经与CsCl 密度梯度离心法比较, 提示两者效果相似, 但本 方法更加经济实用。

     

    Abstract: A method to purify the hepatopancreat ic parvovirus of penaeid shrimp f rom the infected t issue by density gradient centrifug at ion using potassium tart rate-glycerol as subst rate is described in this art icle. Taken f rom the heavily infected shrimp populat ion, the hepatopancrea w as ground thoroughly and treated w ith 1, 1, 2-t richlorot rifuoroethane or chloroform. After the concent rat ion w ith 6% ~ 8% of polyethylene glycol ( PEG) , centrifugat ion through a sucrose cushion of 35% ( w/ w ) was performed. Then, the resulted pellet w as resuspended and centifugalized through a density gradient of potassium tart rate-glycerol prepared according to Ashley ( 1982) . It is indicated from the purif icat ion result that the method is as effect ive as the CsCl density gradient cent rifugation yet less expensive.

     

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