| 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23.3－29.8nm，平均26.3nm，在CsCl中的平均浮力密度为1.360g/cm2；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，40.9kdal和39.8kdal。从而证明，该病毒的确是一种细小病毒。 |
| 关键词: 中国对虾 肝胰腺细小病毒 纯化 鉴定 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金资助项目，39000083号 |
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| PURIFICATION AND CHARACTERIZATION OF THE HEPATOPANCREATIC PARVOVIRUS IN PENAEUS CHINENSIS |
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Xue Qinggang1, Gong Yunhao2, Wang Wenxing1
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1.First Institute of Oceanography, SOA, Qingdao 266003;2.National Animal Quarantine Institute, Ministry of Agriculture, Qingdao 2660032
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| Abstract: |
| During a September, 1991 to end of 1993 study, the hepatopancreatic parvovirus in Penaeus chinensis, named originally as hepatopancreatic parvo-like virus (HPV), was purified from the hepatopancreas of diseased Penaeus chinensis, collected in September 1991 at the Shazikou Shrimp Farm near Qingdao. The virus has been characterized for its biophysical and biochemical properties also. This study showed the virus can justifiably be classified as a member of the Parvoviridae. The purification procedure includes concentrating the virus by differential centrifugation and purifying by density gradient ultracentrifugation through a potassium tartrate-glycerol substrate. The latter was a modified procedure of Ashley’s and the centrifugation time was shortened to 3 hours, 1/5 that of the original method. Ideal purification of the virus was achieved. When negatively stained with neutral phosphotungstic acid solution and examined under electron microscope, the purified virus particles were icosahedrons with average diameter of 26.3nm (23.3 - 29.8 nm). The average diameter of the empty capsid measured 23.2 nm. After extraction from the purified preparation, the viral nucleic acid was treated with DNase, RNase and Nuclease S1 before electrophoresis in agarose gel. The authors finding that the genome nucleic acid of the virus was sensitive to DNase and Nuclease S1 but resistant to RNase. So, it indicates that the viral genome was a single strand DNA molecule about 4.3 kbp long. On SDS-PAGE, the virus showed 4 construction polypeptides, namely VPl, VP2. VP3 and VP4, with molecular weight of about 86.1kdal, 58.2 kdal, 40.9kdal and 39.8 kdal respectively. The average buoyant density of the virus particles was 1.360 g/cm3 in CsCl. All the above characteristics are similar to those of the Parvoviridae. |
| Key words: Penaeus chinensis, Hepatopancreatic parvovirus, Purification characterization |
