Heterologous expression of maize (Zea mays L.) Mir1 cysteine proteinase in eukaryotic and prokaryotic expression systems.
Several heterologous expression systems were tested for their ability to express a unique maize cysteine proteinase Mir1. A baculovirus-based expression system using Trichoplusia ni larvae as host resulted in the expression of Mir1 that was correctly processed and exhibited proteinase activity. Expression in Escherichia coli resulted in accumulation of Mir1, but it had limited solubility and enzymatic activity. Large quantities of Mir1 were produced when Pichia pastoris was used as the host, but the enzyme was insoluble and inactive.
Pechan
T
T
Department of Biochemistry and Molecular Biology, Mississippi State University, MS 39762, USA.
Ma
P W K
PW
Luthe
D S
DS
D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005780:Gelatin / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006458:Hemolymph / Q000378:metabolism; D007814:Larva / Q000235:genetics; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics; D009479:Neuropeptides / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D010449:Peptide Mapping; D010843:Pichia / Q000235:genetics; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012995:Solubility; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D018411:Spodoptera; D003313:Zea mays / Q000201:enzymology* / Q000235:genetics
D009479:Neuropeptides; D010940:Plant Proteins; D011994:Recombinant Proteins; C075028:bombyxins; D005780:Gelatin; D003546:Cysteine Endopeptidases