中国对虾肝胰腺细小病毒的纯化与鉴定  

PURIFICATION AND CHARACTERIZATION OF THE HEPATOPANCREATIC PARVOVIRUS IN PENAEUS CHINENSIS

作　　者：薛清刚[1] 宫云浩[2] 王文兴[1] 

机构地区：[1]国家海洋局第一海洋研究所,青岛266003 [2]农业部动物检疫所,青岛266032

出　　处：《海洋与湖沼》1996年第3期308-313,共6页Oceanologia Et Limnologia Sinica

基　　金：国家自然科学基金!39000083

摘　　要：于1991年9月－1993年底，利用在青岛市沙子口对虾养殖场采集的中国对虾病虾肝胰腺样品，采用一种改进的酒石酸钾－甘油密度梯度离心法，对原先称之为肝胰腺细小样病毒的一种对虾病毒进行纯化，通过电子显微镜观察、提取病毒核酸用不同核酸酶处理并结合电泳等方法，对病毒的基本生物物理和生物化学特性作鉴定。结果表明，用酒石酸钾－甘油密度梯度离心法，可获得良好的病毒纯化效果：纯化后的病毒颗粒显示正二十面体立体对称的特征，颗粒直径为233－29.8nm，平均26．3nm，在CsCl中的平均浮力密度为1．360g/cm3；空的衣壳直径为22－24nm，平均23.2nm；病毒基因组为分子长度相当于4．3kbp的单链DNA，病毒具有4种结构多肽，分子量分别为86．1kdal，58．2kdal，40．gkdal和39.8kdal。从而证明，该病毒的确是一种细小病毒。During a September, 1991 to end of 1993 study, the hepatopancreaticparvovirus in Penaeus chfoensis, named originally as hepatopancreatic parvo-like vi-rus (HPV), was purified from the hepatopancreas of diseased Penaeus chinensis collec-ted in September 1991 at the Shedkou Shrimp Farm near Qingdao. The virus hasbeen characterized for its biophysical and biochendcal properties also. This studyshowed the virus can justifiably be classified as a member of the Parvoviridae. Thepurification procedure includes concentrating the virus by differential centrifugationand purifying by density gradient ultracentrifugation through a potassiumtartrate-glycerol substrate. The latter was a modfied procedure of Ashley's and thecentrifugation time was shortened to 3 hours, 1 /5 that of the original method. Idealpurification of the virus was achieved. When negatively stained with neutralphosphotungstic acid solution and exandned under electron ndcroscope, the purified vi-rus particles were icosahedrons with average diameter of 26.3nm (23.3 - 29.8nm).The average diameter of the empty capsid measured 23.2 nm. After extraction fromthe purified preparation, the viral nucleic acid was treated with DNase, RNase andNuclease S1 before electrophoresis in agarose gel. The authorshnding that the genomenucleic acid of the virus was sensitive to DNase and Nuclease S1 but resistant toRNase. So, it indicates that the viral genome was a single strand DNA moleculeabout 4.3 kbp long. on SDS-PAGE, the virus showed 4 construction polypeptides,namely VP1, VP2, VP3 and VP4, with molecular weight of about 86.1 kdal, 58.2 kdal,40.9kdal and 39.8kdal respectively. The average buoyant density of the virus particleswas 1.360 g /cm3 in CsCl. All the above characteristics are sindlar to those of theParvoviridae.

关 键 词：中国对虾 肝胰腺 病毒 纯化 鉴定 

分 类 号：S945.26[农业科学—水产养殖]