Sf9 insect cells

However, various assays utilizing nonstandardized sources of HDAC protein preparations and substrates appear to generate large variations in the resulting data. As an example. Table 6.1 summarizes the observed variations in the recently reported in vitro data for the effects of SAHA on individual H DAC isoforms [14,19-23]. HDAC isoforms used for in vitro screening assays have been expressed in Escherichia coli (HDACl, 3), Picchia, SF9 insect cells (HDACl-11) or mammalian cells (recombinant HDACl, 3) these sources may lead to differential enzyme activities. 

Besides the currently used constant temperature mode it has been reported that temperature oscillation can enhance cell viability of Sf9 insect cells and ba-culovirus production of occlusion bodies (OB) and extracellular virus (ECV) compared with constant temperature in stationary culture and suspension culture, with the optimal oscillation range at 24-28°C [82]. As a curiosity Pham et al. found that, by raising the infection temperature to 30 °C, they more than doubled the protein productivity of human interleukin-2 (IL-2), in insect larvae, Trichoplusia ni [83]. 

Richardson, M., and Robishaw, J. D. (1999). The a -adrenergic receptor discriminates between G heterotrimers of different fly subunit composition in Sf9 insect cell membranes. / Biol. Chem. 274, 13525—13533. 

Zhang, P-F., Klutch, M., Muller, J., Marcus-Sekura, C. J. Susceptibility of the Sf9 insect cell line to infection with adventitious viruses. Biologicals 22 205-213 (1994). 

Two different protein phosphatases were used the one from Upstate Biotechnology (New York, USA), from human red blood cells, and the one from GTP Technology (Toulouse, France), isolated from SF9 insect cells infected by baculovirus. The enzymatic activity of these two enzymes towards several substrates was investigated by cyclic voltammetry and steady-state chronoamperometry (see experimental details in Refs. [86,87]). First, commercial substrates were tested. Ascorbic acid 2-phosphate and phenyl phosphate were not recognised by the protein... 

Analysis of the primary protein structure of the human 5-HT1B receptor reveals a putative site for palmitoylation, i.e., a cysteine residue located in the short carboxyl tail of the receptor (141). A recombinant c-myc epitope-tagged 5-HT1B receptor was expressed in Sf9 insect cells and palmitoylation of the receptor was demonstrated by metabolic labeling of the cells with [3H]palmitic acid. 

Nebigil CG, et al. Agonist-induced desensitization and phosphorylation of human 5-HT1A receptor expressed in Sf9 insect cells. Biochemistry 1995 34(37) 11,954-11,962. 

Deparis V, Durrieu C, Schweizer M, Marc I, Goergen Jl, Chevalot I, Marc A (2003), Promoting effect of rapeseed proteins and peptides on Sf9 insect cell growth, Cytotechnology 42 75-85. 

Numerous shuttle vectors are available that facilitate convenient transfer from bacteria or mammalian cells. Proteins such as HIV Gp 160, carcinoembryonic antigen, and a form of influenza vaccine have successfully been expressed in baculovirus systems [9]. Potential disadvantages of this system include inappropriate posttranslational modifications, use of insect rather than mammalian cells, and characterization of both the baculovirus and SF9 insect cells used. 

Kunz BC, Muczynski KA, Welsh CF, et al. (1993) Characterization of recombinant and endogenous ADP-ribosylation factors synthesized in Sf9 insect cells. In Biochemistry 32 6643-6648. 

As an alternative, for the recombinant expression of eucaryotic proteins, selectively labeled amino acids can be introduced by the baculovirus expression system in Spodoptera frugiperda (Sf9) insect cells at reasonable costs as shown [28]. Recently, the uniform isotope labeling of the Abl kinase using baculovirus-infected insect cells has been reported [29],... 

Baculovirus infected Spodopterafrugiperda (Sf9) insect cells represent an appropriate expression system for production of membrane proteins and complexes in many applications including biophysical studies and structure determination. The Bac-to-Bac Baculovirus Expression System (Invitrogen) is used to generate bacmids and baculovirus. 

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