Production of recombinant polyhedra containing Cry1Ac fusion protein in insect cell lines.
Insect cell lines and the control of infection for obtaining the maximum amount of polyhedrin-CrylAc-polyhedrin fusion protein from Bactrus in monolayer and suspension culture systems were tested. Growth rates of the Trichoplusia ni (High-Five) cell line in both culture systems were better than the other insect cell lines, Spodoptera fiugiferda (Sf-9, Sf-21), Trichoplusia ni (Tn5), and Spodoptera exigua (Se301). The expression of the fusion protein in a monolayer culture showed that Se301 cells were 2.3-4.8 times more productive on a per cell basis than the other cell lines. However, in suspension culture, only High-Five cells were productive. High-Five cells infected with Bactrus at a multiplicity of infection (MOI) of 5 and a cell density of 3.0 x 10(5) cells per ml were more productive than the other infection condition in a suspension culture suitable for a large-scale production of baculovirus. In conclusion, for the large-scale production of Bactrus in vitro, High-Five cells showing good growth and high productivity are suitable.
Kim
Jae Su
JS
School of Agricultural Biotechnology, College of Agriculture & Life Science, Seoul National University, Seoul 151-742, Korea.
Choi
Jae Young
JY
Roh
Jong Yul
JY
Lee
Han Young
HY
Jang
Seung Sik
SS
Je
Yeon Ho
YH
D000818:Animals; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics; D001427:Bacterial Toxins / Q000096:biosynthesis* / Q000235:genetics; D016367:Baculoviridae / Q000235:genetics / Q000254:growth & development; D002460:Cell Line; D004731:Endotoxins / Q000096:biosynthesis* / Q000235:genetics; D006460:Hemolysin Proteins / Q000096:biosynthesis* / Q000235:genetics; D007915:Lepidoptera; D000077183:Occlusion Body Matrix Proteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D018411:Spodoptera; D015678:Viral Structural Proteins / Q000096:biosynthesis* / Q000235:genetics
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D000077183:Occlusion Body Matrix Proteins; D011993:Recombinant Fusion Proteins; D015678:Viral Structural Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; C009829:polyhedrin protein, Nucleopolyhedrovirus