Recombinant baculovirus

Narum, D. L., Welling, G. W., and Thomas, A. W., Ion-exchange-immuno-affinity purification of a recombinant baculovirus Plasmodium falciparum apical membrane antigen, PF83/AMA-1, /. Chromatogr. A, 657, 357, 1993. 

Assenga, S.P, You, M., Shy, C.H. et al. (2006) The use of a recombinant baculovirus expressing a chitinase from the hard tick Haemaphysalis longicornis and its potential application as a bioacaricide for tick control. Parasitology Research, 98 (2), 111-118. 

Procedures for obtaining recombinant Baculovirus using linearized BaculoGold DNA are simple... 

Protocol 1.7 Cotransfection of insect cells to produce recombinant Baculovirus... 

Sf9 cell culture at 1-2 x 10 cells/ml 1 pig linearized BaculoGold DNA (BD Blosclences) Recombinant Baculovirus transfer vector containing the Insert... 

Mix 0.5 xg linearized BaculoGold DNA with 2-5 xg recombinant Baculovirus transfer vector containing the insert and allow the mixture to sit at room temperature for 5 min. 

Protocol 1.9 Amplification of the recombinant Baculovirus virus stock... 

Kitts, R A. and Possee, R. D. (1993). A method for producing recombinant baculovirus expression vectors at high frequency. Biotechniques 14,810-817. 

The DDI fluorescent inhibition screen is a high-throughput screen, primarily as a result of reduced analysis time, and can range from a few hundred compounds per week to a thousand per week. The assay uses recombinant microsomes prepared from insect cells infected with recombinant baculovirus containing a human CYP enzyme and individual fluorogenic substrates. In this instance the substrates are not specific and hence cannot be used in a mixed enzyme system such as HLM. In addition, this assay is more prone than the conventional inhibition screen (Section 8.3.2) to NCE interferences within the assay (i.e., inherently fluorescent NCEs, fluorescent quenching by the NCE). 

After the BmN cells have started to float, the cell culmre fluid containing the recombinant baculovirus is harvested for silkworm infection and stored at 4°C or -80°C see Note 14). 

The recombinant baculovirus solution is diluted to 50 times with distilled water. 

Hota-Mitchell, S., Siddiqui, A.A., Dekaban, C.A., Smith, J., Tognon, C. and Podesta, R.B. (1997) Protection against Schistosoma mansoni infection with a recombinant baculovirus-expressed subunit of calpain. Vaccine 15, 1631-1640. 

Kawauchi, Y., J. Takasaki, and Y.M. Matsuura. 1994. Preparation and characterization of human rheumatoid arthritic synovial fluid phospholipase A2 produced by recombinant baculovirus-infected insect cells. J Biochem 116 82. 

Germann UA, Willingham MC, Pastan I, et al. Expression of the human multidrug transporter in insect cells by a recombinant baculovirus. Biochemistry 1990 29 (9) 2295-2303. 

Condreay JP, Witherspoon SM, Clay WC, Kost TA (1999), Transient and stable gene expression in mammalian cells transduced with a recombinant baculovirus vector, Proc. Natl Acad. Sci. USA 96 127-132. 

Merrihew RV, Clay WC, Condreay JP, Witherspoon SM, Dallas WS, Kost TA (2001), Chromosomal integration of transduced recombinant baculovirus DNA in mammalian cells, J. Virol. 75 903-909. 

Kamen AA, Bedard C, Tom R, Perret S, Jardin B (1996), On-line monitoring of respiration in recombinant-baculovirus infected and uninfected insect cell bioreactor cultures, Biotechnol. Bioeng. 50 36-48. 

French TJ, Roy P (1990), Synthesis of bluetongue virus (BTV) core-like particles by a recombinant baculovirus expressing the two major structural core proteins of BTV, J. Virol. 64 1530-1536. 

Johnson TM, Li JK (1991) Heterologous expression and characterization of an active lignin peroxidase from Phanerochaete chrysosporium using recombinant baculovirus. Arch Biochem Biophys 291 371-378... 

Recombinant baculoviruses have been used as vectors to express heterologous proteins in cultured insect cells (3). A number of unique features distinguish the baculovirus expression system from other expression systems ... 

Although baculovirus gives a very restricted host range, wild-type and recombinant baculovirus should be treated as potential biohazards. All materials must be either detachment or disinfected before disposal. 

Kitts, P. A. (1995) Production of recombinant baculoviruses using linearized viral DNA. Methods Mol. Biol. 39, 129-142. 

Luckow, V. A., Lee, S. C., Barry, G. F., and Olins, P. O. (1993) Efficient generation of infectious recombinant baculoviruses by site-specific transposon-mediated insertion of foreign genes into a baculovirus genome propagated in Escherichia coli. J. Virol. 67,4566-4579. 

An interesting and more recent expression system for the development of cell-based assays is based on BacMams. These recombinant baculoviruses containing mammalian cell-active expression cassettes seem to be an efficient strategy to speed up assay development. These viruses are produced in insect cells and transiently express but do not replicate in transduced mammalian cells. The expression level can be well controlled by titrating the amount of virus. Highly reproducible transient expression levels, which are a prerequisite to use transient transfection for HTS, might thus be an attractive alternative for some approaches. 

Simplifications caused by recent advances in technology of producing recombinant baculoviruses combined with the ability to infect simultaneously with several different recombinant viruses allows for the production and analysis of multisubunit proteins. 

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