Overview
We have built 3 new Basic Parts and 3 Composite Parts in total. Three Composite Parts are used to express FUCO1,FUCO2 and FUCO3 respectively.
Basic Parts
| Description | Type | Name |
|---|---|---|
| CYC1 | Terminator | BBa_K3812006 |
| BW25113 | RBS | BBa_K3812011 |
| 6xHis-tag | Tag | BBa_K3812012 |
| Paox1 | Promoter | BBa_K3812005 |
| pSB1C3 | Plasmid | BBa_K3812014 |
| FUCO1 | Coding | BBa_K4688002 |
| FUCO2 | Coding | BBa_K4688003 |
| FUCO3 | Coding | BBa_K4688004 |
Composite Parts
| Description | Type | Name |
|---|---|---|
| FUCO1-GFP-expression | Composite | BBa_K4688005 |
| FUCO2-GFP-expression | Composite | BBa_K4688006 |
| FUCO3-GFP-expression | Composite | BBa_K4688007 |
Validation
The FUCO-encoded fucosidases of P. tricornutum was cloned by overlap PCR. It is responsible for hydrolyzing the alpha-1,6-linked fucose joined to the reducing-end N-acetylglucosamine of the carbohydrate moieties of glycoproteins and thus cleaving the nonreducing terminal-L-fucose from these glycoconjugates.
Constructed recombinant vector, transferred to E. coli and tested.
