Abstract
The group B entomopoxvirus (EPV) from Amsacta moorei (AmEPV) productively infects only insect cells. A series of AmEPV-lacZ recombinants was constructed in which the lacZ gene was regulated by either late (the AmEPV spheroidin or the cowpox virus A-type inclusion [ATI]) or early (the AmEPV esp [early strong promoter; derived from a 42-kDa AmEPV protein] or the Melolontha melolontha EPV fusolin, fus) virus promoters. When the AmEPV recombinants were used to infect vertebrate cells, beta-galactosidase expression occurred (in >30% of the cells) when lacZ was regulated by either the fus or esp early promoters but not when lacZ was regulated by the late promoters (spheroidin or ATI). Therefore, AmEPV enters vertebrate cells and undergoes at least a partial uncoating and early, but not late, viral genes are expressed. Neither viral DNA synthesis nor cytopathic effects were observed under any infection conditions. When an AmEPV recombinant virus containing the Aequorea victoria green fluorescent protein gene (gfp) under the control of the esp promoter was used to infect vertebrate cells at a low multiplicity of infection, single fluorescent cells resulted, which continued to divide over a period of several days, ultimately forming fluorescent cell clusters, suggesting that vertebrate cells survive the infection and continue to grow. Therefore, AmEPV may prove to be a highly efficient, nontoxic method of gene delivery into vertebrate cells for transient gene expression.
Full Text
The Full Text of this article is available as a PDF (1.5 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Arif B. M. Recent advances in the molecular biology of entomopoxviruses. J Gen Virol. 1995 Jan;76(Pt 1):1–13. doi: 10.1099/0022-1317-76-1-1. [DOI] [PubMed] [Google Scholar]
- Boyce F. M., Bucher N. L. Baculovirus-mediated gene transfer into mammalian cells. Proc Natl Acad Sci U S A. 1996 Mar 19;93(6):2348–2352. doi: 10.1073/pnas.93.6.2348. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Chalfie M., Tu Y., Euskirchen G., Ward W. W., Prasher D. C. Green fluorescent protein as a marker for gene expression. Science. 1994 Feb 11;263(5148):802–805. doi: 10.1126/science.8303295. [DOI] [PubMed] [Google Scholar]
- Christen L., Seto J., Niles E. G. Superinfection exclusion of vaccinia virus in virus-infected cell cultures. Virology. 1990 Jan;174(1):35–42. doi: 10.1016/0042-6822(90)90051-R. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Condit R. C., Motyczka A. Isolation and preliminary characterization of temperature-sensitive mutants of vaccinia virus. Virology. 1981 Aug;113(1):224–241. doi: 10.1016/0042-6822(81)90150-1. [DOI] [PubMed] [Google Scholar]
- Dall D., Sriskantha A., Vera A., Lai-Fook J., Symonds T. A gene encoding a highly expressed spindle body protein of Heliothis armigera entomopoxvirus. J Gen Virol. 1993 Sep;74(Pt 9):1811–1818. doi: 10.1099/0022-1317-74-9-1811. [DOI] [PubMed] [Google Scholar]
- Fischer L., Le Gros F. X., Mason P. W., Paoletti E. A recombinant canarypox virus protects rabbits against a lethal rabbit hemorrhagic disease virus (RHDV) challenge. Vaccine. 1997 Jan;15(1):90–96. doi: 10.1016/s0264-410x(96)00102-8. [DOI] [PubMed] [Google Scholar]
- Gauthier L., Cousserans F., Veyrunes J. C., Bergoin M. The Melolontha melolontha entomopoxvirus (MmEPV) fusolin is related to the fusolins of lepidopteran EPVs and to the 37K baculovirus glycoprotein. Virology. 1995 Apr 20;208(2):427–436. doi: 10.1006/viro.1995.1173. [DOI] [PubMed] [Google Scholar]
- Gruidl M. E., Hall R. L., Moyer R. W. Mapping and molecular characterization of a functional thymidine kinase from Amsacta moorei entomopoxvirus. Virology. 1992 Feb;186(2):507–516. doi: 10.1016/0042-6822(92)90016-i. [DOI] [PubMed] [Google Scholar]
- Hall R. L., Li Y., Feller J., Moyer R. W. The Amsacta moorei entomopoxvirus spheroidin gene is improperly transcribed in vertebrate poxviruses. Virology. 1996 Oct 15;224(2):427–436. doi: 10.1006/viro.1996.0549. [DOI] [PubMed] [Google Scholar]
- Hall R. L., Moyer R. W. Identification, cloning, and sequencing of a fragment of Amsacta moorei entomopoxvirus DNA containing the spheroidin gene and three vaccinia virus-related open reading frames. J Virol. 1991 Dec;65(12):6516–6527. doi: 10.1128/jvi.65.12.6516-6527.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hofmann C., Sandig V., Jennings G., Rudolph M., Schlag P., Strauss M. Efficient gene transfer into human hepatocytes by baculovirus vectors. Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10099–10103. doi: 10.1073/pnas.92.22.10099. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kawakita M., Rao G. S., Ritchey J. K., Ornstein D. K., Hudson M. A., Tartaglia J., Paoletti E., Humphrey P. A., Harmon T. J., Ratliff T. L. Effect of canarypox virus (ALVAC)-mediated cytokine expression on murine prostate tumor growth. J Natl Cancer Inst. 1997 Mar 19;89(6):428–436. doi: 10.1093/jnci/89.6.428. [DOI] [PubMed] [Google Scholar]
- Langridge W. H. Detection of Amsacta moorei entomopoxvirus and vaccinia virus proteins in cell cultures restrictive for poxvirus multiplication. J Invertebr Pathol. 1983 Jul;42(1):77–82. doi: 10.1016/0022-2011(83)90204-5. [DOI] [PubMed] [Google Scholar]
- Muldoon R. R., Levy J. P., Kain S. R., Kitts P. A., Link C. J., Jr Tracking and quantitation of retroviral-mediated transfer using a completely humanized, red-shifted green fluorescent protein gene. Biotechniques. 1997 Jan;22(1):162–167. doi: 10.2144/97221rr03. [DOI] [PubMed] [Google Scholar]
- Paoletti E. Applications of pox virus vectors to vaccination: an update. Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11349–11353. doi: 10.1073/pnas.93.21.11349. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pedley C. B., Cooper R. J. The assay, purification and properties of vaccinia virus-induced uncoating protein. J Gen Virol. 1987 Apr;68(Pt 4):1021–1028. doi: 10.1099/0022-1317-68-4-1021. [DOI] [PubMed] [Google Scholar]
- Smith G. L., Moss B. Infectious poxvirus vectors have capacity for at least 25 000 base pairs of foreign DNA. Gene. 1983 Nov;25(1):21–28. doi: 10.1016/0378-1119(83)90163-4. [DOI] [PubMed] [Google Scholar]
- Taylor J., Paoletti E. Fowlpox virus as a vector in non-avian species. Vaccine. 1988 Dec;6(6):466–468. doi: 10.1016/0264-410x(88)90091-6. [DOI] [PubMed] [Google Scholar]
- Taylor J., Weinberg R., Languet B., Desmettre P., Paoletti E. Recombinant fowlpox virus inducing protective immunity in non-avian species. Vaccine. 1988 Dec;6(6):497–503. doi: 10.1016/0264-410x(88)90100-4. [DOI] [PubMed] [Google Scholar]
- Tine J. A., Lanar D. E., Smith D. M., Wellde B. T., Schultheiss P., Ware L. A., Kauffman E. B., Wirtz R. A., De Taisne C., Hui G. S. NYVAC-Pf7: a poxvirus-vectored, multiantigen, multistage vaccine candidate for Plasmodium falciparum malaria. Infect Immun. 1996 Sep;64(9):3833–3844. doi: 10.1128/iai.64.9.3833-3844.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Winter J., Hall R. L., Moyer R. W. The effect of inhibitors on the growth of the entomopoxvirus from Amsacta moorei in Lymantria dispar (gypsy moth) cells. Virology. 1995 Aug 20;211(2):462–473. doi: 10.1006/viro.1995.1428. [DOI] [PubMed] [Google Scholar]
- Yuen L., Dionne J., Arif B., Richardson C. Identification and sequencing of the spheroidin gene of Choristoneura biennis entomopoxvirus. Virology. 1990 Apr;175(2):427–433. doi: 10.1016/0042-6822(90)90427-s. [DOI] [PubMed] [Google Scholar]
- Zolotukhin S., Potter M., Hauswirth W. W., Guy J., Muzyczka N. A "humanized" green fluorescent protein cDNA adapted for high-level expression in mammalian cells. J Virol. 1996 Jul;70(7):4646–4654. doi: 10.1128/jvi.70.7.4646-4654.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]