Signal sequence is still required in genes downstream of "autocleaving" 2A peptide for secretary or membrane-anchored expression.
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.
Yan
Jun
J
Department 4, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing 400042, People's Republic of China.
Wang
Hongjun
H
Xu
Qian
Q
Jain
Nidhi
N
Toxavidis
Vasilis
V
Tigges
John
J
Yang
Hailin
H
Yue
Guohua
G
Gao
Wenda
W
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
D007074:Immunoglobulin G; D007376:Interleukin-2; D008565:Membrane Proteins; D010455:Peptides; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D018800:Thy-1 Antigens