title: Signal sequence is still required in genes downstream of "autocleaving" 2A peptide for secretary or membrane-anchored expression.
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.
authors: Department 4, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing 400042, People's Republic of China.
authors: Jun
authors: J
authors: Yan
authors: Hongjun
authors: H
authors: Wang
authors: Qian
authors: Q
authors: Xu
authors: Nidhi
authors: N
authors: Jain
authors: Vasilis
authors: V
authors: Toxavidis
authors: John
authors: J
authors: Tigges
authors: Hailin
authors: H
authors: Yang
authors: Guohua
authors: G
authors: Yue
authors: Wenda
authors: W
authors: Gao
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000378:metabolism; D007376:Interleukin-2 / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism*; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D055571:Secretory Pathway; D018800:Thy-1 Antigens / Q000378:metabolism
chemical_list: D007074:Immunoglobulin G; D007376:Interleukin-2; D008565:Membrane Proteins; D010455:Peptides; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D018800:Thy-1 Antigens