Abstract
2A Peptide sequences are now being widely used to construct multicistronic expression vectors. It is suggested that when only the first 2A-linked protein bears a signal sequence, the signal-less protein(s) downstream of 2A can also be translocated into the mammalian endoplasmic reticulum system through a "slipstreaming" mechanism. By using flow cytometry and cell surface CD90 as a localization indicator, we show here that slipstreaming translocation does not occur in mammalian cells; that is, the second protein downstream of 2A still requires signal sequence for secretary or membrane-anchored expression.
Copyright 2009 Elsevier Inc. All rights reserved.
Publication types
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Research Support, Non-U.S. Gov't
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Technical Report
MeSH terms
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Animals
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CHO Cells
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Cricetinae
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Cricetulus
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Endoplasmic Reticulum / metabolism
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Humans
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Immunoglobulin G / genetics
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Immunoglobulin G / metabolism
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Interleukin-2 / genetics
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Interleukin-2 / metabolism
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Membrane Proteins / chemistry
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Membrane Proteins / genetics
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Membrane Proteins / metabolism*
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Peptides / metabolism*
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Protein Sorting Signals*
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / metabolism
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Secretory Pathway*
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Thy-1 Antigens / metabolism
Substances
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Immunoglobulin G
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Interleukin-2
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Membrane Proteins
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Peptides
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Protein Sorting Signals
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Recombinant Fusion Proteins
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Thy-1 Antigens