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Expression of Bombyx mori nucleopolyhedrovirus ORF76 in permissive and non-permissive cell lines by a novel Bac-to-Bac-BmNPV baculovirus expression system

100%

Su W. J., Wu Y., Wu H.-L., Zhu S.-Y., Wang W.-B.

Polish Journal of Microbiology

2008

tom 57

nr 4

Open reading frame 76 of Bombyx mori nucleopolyhedrovirus (BmNPV), designated as Bm76, is a gene whose function is completely unknown. With EGFP fused to the 3' terminal of Bm76 as the reporter gene and BmNPV bacmid as the expression vector, a recombinant bacmid was successfully constructed expressing Bm76-EGFP fusion protein under the control of polyhedrin promoter in Bombyx mori cells (Bm cells), BmNPV's permissive cell line, laying the foundation for rescue experiment of Bm76 deletion mutant. Moreover, the supernatant from Bm cells transfected with the recombinant bacmid was used to infect Trichoplusia Ni cells (Tn cells), BmNPV's non-permissive cell line. Unexpectedly, the expression of Bm76-EGFP fusion protein in some Tn cells was detected, implying that viral DNA was replicated in these cells. The causes are being studied for the inability of BmNPV to produce enough viable budded viruses in Tn cells despite of viral DNA replication.

The effect of a small conotoxin-like ctx gene from Autographa californica nuclear polyhedrosis virus (AcMNPV) on insect hemolymph melanization

86%

Cao Q., Zhu S.-Y., Wu Y., Liu Y., Zhu J., Wang W.

Polish Journal of Microbiology

2012

tom 61

nr 3

The conotoxin-like (ctx) gene encodes a small cysteine-rich polypeptide in various baculoviruses. Previous research has demonstrated that the product of the ctx gene could be purified from insect cells infected by Autographa californica nuclear polyhedrosis virus (AcMNPV), but its function was unknown. In this paper, we compared the conserved cysteine motif structure (CX3GX2CX5CCX3CX6C) of the ctx gene in baculoviruses and generated recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV) with the BmNPV bacmid system. !e recombinant BmNPV contained the ctx gene from AcMNPV or a fusion gene of ctx with eGFP, respectively. Fluorescence in CTX-eGFPpositive cells was mainly observed on the cell membrane. To gain insight into CTX function, two methods were used to elucidate the affect CTX had on hemolymph melanization in vivo and in vitro in insect larvae and pupae. The results indicated that CTX abrogates hemolymph melanization; however, the mechanisms require further evaluation.

Ograniczanie wyników

2 Polish Journal of Microbiology

2 Wu Y.

2 Zhu S.-Y.

1 Cao Q.

1 Liu Y.

1 Su W. J.

1 Wang W.

1 Wang W.-B.

1 Wu H.-L.

1 Zhu J.

1 2012

1 2008

Wyniki wyszukiwania

Expression of Bombyx mori nucleopolyhedrovirus ORF76 in permissive and non-permissive cell lines by a novel Bac-to-Bac-BmNPV baculovirus expression system

The effect of a small conotoxin-like ctx gene from Autographa californica nuclear polyhedrosis virus (AcMNPV) on insect hemolymph melanization