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Signal sequence is still required in genes downstream of “autocleaving” 2A peptide for secretary or membrane-anchored expression |
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| Authors: | Jun Yan Qian Xu Vasilis Toxavidis Hailin Yang Wenda Gao |
| Institution: | a Department 4, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing 400042, People’s Republic of China b Department of Surgery, Transplant Institute, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA c Brandeis University, Waltham, MA 02453, USA d Department of Medicine, Transplant Institute, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA e Antagen Biosciences, Boston, MA 02215, USA |
| 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. |
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