EC Number   |
Reference |
|---|
   2.4.1.144 | - |
638228, 672706 |
| 2.4.1.144 | amplified for cloning into pENTR-D-Topo vector, cloned gene inserted into the virus expression vector, pBABE-puro. GnT-III and GnT-V constructs transfected into Phoenix-Ampho cells |
704468 |
| 2.4.1.144 | cloning from genetic library, DNA sequence determination, functional overexpression in COS-1 or HeLa-cells, transient transfection |
638214 |
| 2.4.1.144 | cloning of cDNA by RT-PCR |
638227 |
| 2.4.1.144 | construction of genes for expression of native rat GnTIII, human ManII, Arabidopsis thaliana ManII and for the expression of the catalytic domains of rat GnTIII and human ManII fused to the N-terminal region of Arabidopsis thaliana ManII. Binary plasmids for high-level expression of rat GnTIII (pAX67), chimeric rat GnTIIIA.th. (replaced native localization domain with the cytoplasmic tail, transmembrane, and stem region of Arabidopsis thaliana mannosidase II) (pAX70), co-expression of rat GnTIII together with human ManII (pAX73) or Arabidopsis thaliana ManII (pAX100), co-expression of GnTIIIA.th. together with ManIIA.th. (pAX74), and co-expression of GnTIIIA.th. and Arabidopsis thaliana ManII (pAX101). Expression of the transgenes is driven by a chimeric promoter assembled from regulatory elements from mannopine and octopine synthase genes. Expression cassettes comprising the CaMV 35S promoter, GnTIII or ManII and the nos pA, being pSK103 (ManII), pSK124 (ManII-Arabidopsis thaliana), pAX63 (GnTIII-Arabidopsis thaliana) and pAX68 (GnTIII) serve as starting point for further expression constructs. The cassette comprising the CaMV 35 promoter, plant relocalized GnTIII and polyadenylation signal is excised from pAX68 using SbfI, EcoRI and inserted into pAX36 generating pAX90. The construct comprising the rat GnTIII obtained by replacing the XbaI, AflII fragment from pAX90 with the fragment isolated from pAX63, generating pAX91. Mutated fragment from pCLF40 inserted into pAX69 using AvaI, StuI, generating pAX104. The binary expression plasmids for inactive GnTIII and GnTIIIA.th. under the control of the UAS123mas promoter generated by replacing the StuI, EcoRI fragment from pAX104 with that in pAX67 (generating pAX105) and pAX70 (generating pAX106), respectively. The mutated fragment from pCLF40 inserted into pAX91 using Eam1105I, StuI, generating the expression plasmid for inactive GnTIII under the control of the CaMV 35S promoter (pAX108). The exchange of the fragment containing the Arabidopsis thaliana Golgi localization domain from pAX90 in pAX108 using SbfI, StuI yields an expression plasmid for inactive GnTIIIA.th. under the control of the CaMV 35S promoter. Plasmids transferred into Agrobacterium tumefaciens LBA4404 for transformation into tobacco BY-2 cells |
704773 |
| 2.4.1.144 | construction of transgenic mice, disruption of certain functions of apolipoprotein B leading to generation of a aftty liver |
638223, 638228 |
| 2.4.1.144 | enzyme overexpression in HeLa-S3, CHO-K1, Pro-5, and MDA-MB-231 cells using vector CSIV-TRE-RfA-CMV-KT, cloning in HEK-293T cells |
736507 |
| 2.4.1.144 | expressed in Spodoptera frugiperda Sf21 cells |
722343 |
| 2.4.1.144 | expressed in transgenic mice |
705381 |
| 2.4.1.144 | expression in erythroleukemia cell line K562, leading to increased resistance to lysis by natural killer cells and enhanced spleen colonization ability |
638223, 638228 |
