分离草虾杆状病毒包涵体的一种新方法

A new approach to purify occlusion bodies of Penaeus monodon-type baculovirus

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／dm3Tris－HCIpH8．0，50mmol／dm3EDTA，0．1mol／dm3NaCl，0.5mmol／dm3PMSF）破碎，差迷离心，再经30％～70％蔗糖浓度梯度高速离心，可得到合包涵体的区带。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。

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.