Table 1.
Glycoenzyme constructs used in this study.
| Glycoenzyme* | ie1 plasmid (reference) | 39K plasmid | Method used for 39K plasmid construction |
|---|---|---|---|
| MGAT1 | pIE1GlcNAcTI (Hollister et al., 2002) | p39KHRGlcNAcTI | 2.7 kb NotI-HindIII fragment from pIE1GlcNAcTI subcloned into NotI-HindII sites of p39K-SEAP |
| MGAT2 | pIE1GlcNAcTII (Hollister et al., 2002) | p39KHRGlcNAcTII | 2.6 kb BglII-HindIII fragment from pIE1GlcNAcTII subcloned into BglII-HindIII sites of p39K- SEAP |
| B4GALT1 | pIE1HR3HyGalT(F282) (Geisler, in prep) | p39KHR3HyGalT(F282) | 1.0 kb BglII-NruI fragment from pIE1HR3HyGalT(F282) subcloned into BglII-PmeI sites of p39K- SEAP |
| GNPE | pIE1HREcGNPE (Geisler and Jarvis, 2012a) | p39KHREcGNPE | 2.0 kb BglII-HindIII fragment from pIE1EcGNPE subcloned into BglII-HindIII sites of p39K-SEAP |
| SAS | pIE1HRMmSAS (Geisler and Jarvis, 2012a) | p39KHRMmSAS | 2.4 kb NotI-HindIII fragment from pIE1HRMmSAS subcloned into NotI-HindIII sites of p39K-SEAP |
| CMAS | pIE1HRMmCSAS (Geisler and Jarvis, 2012a) | p39KHRMmCSAS | 2.1 kb BamHI-XbaI fragment from pIE1MmCSAS subcloned into BglII-XbaI sites of p39K-SEAP |
| CSAT | pIE1-hCSAT (Mabashi-Asazuma et al., 2013) | p39K-hCSAT | 2.3 kb BglII-HindIII fragment from pIE1-hCSAT subcloned into BglII-HindII sites of p39K-SEAP |
| ST3GAL4 | pIE1-hST3GAL4b (Mabashi, in prep) | p39K-hST3GAL4b | 1.8 kb BamHI-XbaI fragment from pIE1-hST3GAL4b subcloned into BglII-XbaI sites of p39K-SEAP |
| ST6GAL1 | pIE1HRHyST6Δcys (Geisler, in prep) | p39KHRHyST6Δcys | 1.9 kb SacII(3′ overhangs removed with T4 DNA Polymerase)-XbaI fragment from pIE1HRHyST6Δcys subcloned into NotI(5′ overhangs filled with T4 DNA Polymerase)- XbaI sites of p39K-SEAP |
HUGO nomenclature, except CSAT, whose HUGO symbol is SLC35A1