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中国对虾杆状病毒基因克隆及探针制备与检测

Partial Cloning of the Genome of Non occluded Baculovirus from Penaeus chinensis and Preparing the Probe for Detection

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【作者】 石正丽; 黄灿华; 陈棣华; J.R.Bonami; S.Durand;

【Author】 Shi Zhengli\ \ Huang Canhua\ \ Chen Dihua (Wuhan Institute of Virology, Academia Sinica,Wuhan 430071) J.R.Bonami\ \ S.Durand (UMR219,CNRS IFREMER,UMII,Place E.Bataillon, 34095 Montpellier,Cedex 5,France)

【机构】 中国科学院武汉病毒研究所;

【摘要】 从收集的中国对虾(Penaeuschinensis)病虾样品中分离到一种杆状病毒,经负染在电镜下观察,完整病毒粒子大小为110×280~320nm。病毒核酸经EcoRI酶切,克隆到pUC18质粒上,经筛选得到两个克隆L46、M13,克隆片段分别为2.14kb和3.58kb。将这两个基因片段和对虾白斑综合征杆状病毒(WhiteSpotSyndromeBaculovirus,简称WSSV)基因克隆片段A26制备探针,共同用点杂交法对我国沿海地区的病虾样品进行检测,以了解我国沿海地区对虾杆状病毒的分布,并确定中国对虾杆状病毒与WSSV的同源性。

【Abstract】 Viral particles similar to non occluded baculoviruses were isolated from penaeid shrimp Penaeus chinensis in China. The size of enveloped virion is 250-300×110nm.After extraction,the viral DNA was digested by EcoRⅠ and cloned into the pUC 18. Two clones, L46 and M13,containing inserts of 2.14kb and 3.58kb in size respectively, were obtained.The inserts were labeled by Digoxingenin. Diseased shrimp samples issued from different regions of China were investigated by dot blot hybridization with the previously preparing probes L46, M13 and with A26 probe. All the probes reacted with the tested samples showing baculovirus infection. It indicated that baculovirus from P.chinensis is very close to the WSSV.

【关键词】 中国对虾; 杆状病毒; 基因克隆; 核酸探针;
【Key words】 Penaeus chinensis; Baculovirus; Gene cloning; DNA probe;
  • 【文献出处】 中国病毒学 ,VIROLOGICA SINICA , 编辑部邮箱 ,1998年03期
  • 【分类号】Q939.47,
  • 【被引频次】39
  • 【下载频次】137
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