The Study Of Association Of The Replication Of HaSNPV With Nuclear Matrix In Hz Cells

Association of the replication of HaSNPV with nuclear matrix was studied in Hz cells. Nuclear matrices from uninfected Hz cells and those infected with HaSNPV were isolated.By the whole-mount electron microscopy, a subtle network of nuclear matrix was found in Hz cells,and nuclear matrix had no dramtic morphological change in early infection stage, polyhedra appeared to be attached to the network of the nuclear matrix in the very late stage of infection .By thin section not only polyhedra but also virions and nucleocapsids were attached to the network of nuclear matrix. The protein constituentes of nuclear matrix of Hz cells uninfected and infected by HaSNPV at different time were compared by SDS-PAGE, the enhancing protein bands at about 32KD, 3OKD, 29KD were found in the nuclear matrix protein of Hz cells 48 houres post infection(hpi).Because the molecular weight of capsid protein and another structural polypeptide of polyhedra-derived virus (PDV)were respectively 32KD, 3OKD,and the molecular weight of polyhedrin was 29KD,it was thought that the enhancing bands of 32KD, 3OKD in nuclear matrix may be the capsid protein and another structural polypeptide of PDV which were bond to nuclear matrix. The antibody of HaSNPV PDV (anti-PDV) was prepared and purified,and was used to confirm that capsid protein and another structural polypeptide of PDV bond to the nuclear matrix of Hz cells 48hpi.Whether polyhedrin bond to the nuclear matrix of Hz cells 48hpi should be further confirmed by immunology method and Western blot. Indirect gold-immunelabeled colloid electron microscopy were employed to examine if there was actin in nuclear matrix of Hz cells(primary antibody:anti-actin,second antibody: 10-nm gold-labeled IgG). No gold particle was found in the control,but many gold garticles were found in the nuclear matrix tested and their arrangement had no obvious a regulation. Western blot has confirmed that:l.There was actin in nuclear matrix of Hz cells.2. With the infection processing, the amount of actin dropped gradually. It was also confirmed by Western blot that actin did not specially bind to budded virus, polyhedra梔erived virus and their nucleocapsid.