| 中国对虾一种C型杆状病毒的纯化技术及形态特征研究 |
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| 引用本文： | 孔杰,石拓,刘萍,包振民,郭福生,相建海.中国对虾一种C型杆状病毒的纯化技术及形态特征研究[J].海洋与湖沼,1997,28(3):233-237. |
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| 作者姓名： | 孔杰 石拓 刘萍 包振民 郭福生 相建海 |
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| 作者单位： | 中国水产科学研究院黄海水产研究所!青岛，266071，中国水产科学研究院黄海水产研究所!青岛，266071，中国水产科学研究院黄海水产研究所!青岛，266071，青岛海洋大学生命科学院!青岛，266003，农业部动物检疫所!青岛，266071，中国科学院海洋研究所!青岛，266071 |
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| 基金项目： | 国家攀登计划“B”资助!PDB6－6－1 |
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| 摘 要： | 染病中国对虾于１９９５年７月取自青岛崂山对养殖场。为探索对虾杆状病毒分离纯化技术，采用组织超薄切片技术对染病对虾进行电镜观察：被病毒感染的对虾经组织匀浆、差速离心和密度梯度离心，纯化出杆状病毒粒子；纯化的杆状病毒经钨酸钾负染色，于透射电子显微镜下观察形态结构。结果表明，中国对虾中肠组织
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| 关 键 词： | 中国对虾 病毒 纯化 形态特征 虾 |
| 收稿时间： | 1996/11/20 0:00:00 |
| 修稿时间： | 1997/1/10 0:00:00 |
STUDIES ON PURIFICATION TECHNIQUE AND MORPHOLOGY OF A KIND OF TYPE C BACULOVIRUS IN PENAEUS CHINENSIS |
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| Kong Jie,Shi Tuo,Liu Ping,Bao Zhenmin,Guo Fusheng and Xiang Jianhai.STUDIES ON PURIFICATION TECHNIQUE AND MORPHOLOGY OF A KIND OF TYPE C BACULOVIRUS IN PENAEUS CHINENSIS[J].Oceanologia Et Limnologia Sinica,1997,28(3):233-237. |
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| Authors: | Kong Jie Shi Tuo Liu Ping Bao Zhenmin Guo Fusheng and Xiang Jianhai |
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| Institution: | Yellow Sea Fisheries Research Institute, Chinese Acadimy of Fishery Sciences, Qingdao 266071;Yellow Sea Fisheries Research Institute, Chinese Acadimy of Fishery Sciences, Qingdao 266071;Yellow Sea Fisheries Research Institute, Chinese Acadimy of Fishery Sciences, Qingdao 266071;College of Life Sciences,Ocean University of Qingdao,266003;National Animal Quarantine Institute, Ministry of Agriculture, Qingdao 266071;Institute of Oceanology , Chinese Academy of Sciences, Qingdao 266071 |
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| Abstract: | Moribund shrimps, Peneaus chinensis (3 - 5 cm in body length), werecollected during an occurrence of mass mortality at a shrimp farm of Lnoshan, Qingdaoin July 1995. The gill tissue, hepatopancreatic tissue and mid-gut tissue were fixatedfor ultrathin sectioning and the ealstenee of a kind of baculovirus was found in thegill and mid-gut by Hitachi H-7000 transndssion electron microscope. Cytopathicchanges associated with baculovirus infection were hypertrophy of the nucleus andaccumuation of rod-shaped enveloped virions (Plate I:l). To isolate the virus, 100g ofmoribun shrimp were homogenized at 6 000 r/min for 5 ndn with PPB buffer (HuangJie, personal communication) or PPB buffer containing 2% Triton X- 100. The outermembrane of the virions was broken and the virions were purified as nucleocapsids inthe presence of Trition X-100. Pellets of the material homogenated at differentcentrifugation speeds were inspected by electron ndcroscope and baculovirus wasfound to be precipitated at 8000 r /min (Tab. l)- Morphologically intact baculovirusand nucleocapsids could be isolated and purified by centrifugation from 6000r /min tol0 000r /min and density gradient centrifugation on l0%- 50% sugrose at 25 000 r /min from cephalothorax homogenate. The morphology and structure of the virus, stainedby phosphotungstic acid (PTA), were analyzed by transndssion electron ndcroscope'Morphologically intact baculovirus were rod-shaped, singly enveloped -and with along tail-like appendage' The head of whole virions averaged 90- l30 nm in diame-ter and 300 - 370nm in length' The Tail was 2O - 30nm in diameter and 500-700nm in length (Plate 1:2). The nucleocapsid contained l4 helixes with 2 cappedends on both sides and formed bacilliform structure (Plate I:3)' |
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| Keywords: | Peneaus chinensis Virus Purification |
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