Use of the signal peptide of Pisum vicilin to translocate beta-glucuronidase in Nicotiana tabacum.
A hybrid protein system was used for the study of protein transport in plant cells. A nucleotide sequence (vic) encoding a putative signal peptide of 15 amino acid residues, derived from the published aa sequence of one Pisum vicilin, was synthesized and fused in frame to the gus gene encoding a bacterial cytosolic beta-glucuronidase (GUS). When the hybrid vic::gus gene was expressed in tobacco cells using the cauliflower mosaic virus 35S promoter, the hybrid GUS protein was targeted to, and glycosylated inside the rough endoplasmic reticulum. Glycosylation could be blocked with the antibiotic tunicamycin. The study of transient expression in protoplasts showed that extracellular secretion efficiency was low, which may be due to the nature of the GUS protein.
Pang
S Z
SZ
Department of Horticultural Sciences, Cornell University, Geneva, NY 14456.
Rasmussen
J
J
Ye
G N
GN
Sanford
J C
JC
D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport / Q000502:physiology; D015870:Gene Expression / Q000235:genetics; D005966:Glucuronidase / Q000378:metabolism*; D006031:Glycosylation / Q000187:drug effects; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics; D010940:Plant Proteins; D014674:Plant Proteins, Dietary / Q000235:genetics* / Q000378:metabolism; D010944:Plants / Q000235:genetics; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D055314:Seed Storage Proteins; D014026:Nicotiana / Q000378:metabolism*; D014415:Tunicamycin / Q000494:pharmacology
D010940:Plant Proteins; D014674:Plant Proteins, Dietary; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D055314:Seed Storage Proteins; D014415:Tunicamycin; C015575:vicilin protein, plant; D005966:Glucuronidase