| 分离草虾杆状病毒包涵体的一种新方法 |
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| 引用本文： | 杨丰,林斌,等.分离草虾杆状病毒包涵体的一种新方法[J].台湾海峡,1995,14(1):80-82. |
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| 作者姓名： | 杨丰 林斌 |
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| 作者单位： | [1]国家海洋局第三海洋研究所，厦门361005 [2]厦门大学抗癌 |
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| 基金项目： | 厦门市虾病攻关专项基金 |
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| 摘 要： | 本文报道了一种纯化草虾杆状病毒（MBV）包涵体的方法。取出受MBV感染的虾肝胰组织，在TESP溶液中（50mmol/dm^3Tris-HC1 pH8.0,50mmol/dm^3EDTA,0.1mol/dm^3NaC1,0.5mmol/dm^3PMSF）破碎，差速离心，再经30%～70%蔗糖浓度梯度高速离心，可得到含包涵体的区带。电镜下观察到较纯的包涵体，经蛋白酶K降解后，PCR扩增，经琼脂糖凝胶电泳可见清晰的DNA区带。
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| 关 键 词： | 虾病毒 分离包涵体 PCR鉴定 草虾杆状病毒 |
A new approach to purify occlusion bodies of Penaeus monodon-type baculovirus |
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| Yang Feng, Lin Bin, Chen Xifa'' and Xu Xun.A new approach to purify occlusion bodies of Penaeus monodon-type baculovirus[J].Journal of Oceanography In Taiwan Strait,1995,14(1):80-82. |
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| Authors: | Yang Feng Lin Bin Chen Xifa' and Xu Xun |
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| Abstract: | A new approach to purify occlusion bodies (OB) of Penaeus monodon-type baculovirus(MBV) is described. MBV-infected hepatoancrafa were removed from the shrimps andhomogenized in a solution of TESP buffer (50mmol/dm' Tris-HCl pH 8.0, 50mmol/dm3EDTA, 0. 1mol/dm3 NaCl, 0. 5mmol/dm3 PMSF). PMSF (phenylmethyl solfonyl flouride) wasused as a proteinase inhibitor protecting OB from degradation. The suspension was clarifiedin differential cantrifugation and the pellet was further separted in a 30%~ 70% sucrosegradient by high speed centrifugation. Occlusion bodies corresponding to a specific band inthe gradient was collected and observed under an electron microscope. After digestion byproteinase K, the MBV DNA was amplified by PCR and visualized on an agarose gel. |
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| Keywords: | Shrip virus isolation of occlusion bodies MBV PCR |
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