front |1 |2 |3 |4 |5 |6 |7 |8 |9 |10 |11 |12 |13 |14 |15 |16 |17 |18 |19 |20 |21 |22 |23 |24 |25 |26 |27 |28 |29 |30 |31 |32 |33 |34 |review |
The modified
secretory-trap vector incorporates a transmembrane (TM) domain, an internal
ribosome entry site (IRES) and an alkaline phosphatase (PLAP) gene. Only the
situation after insertion of this vector into a gene encoding a protein with
a hydrophobic leader sequence (S) is shown. A bicistronic transcript is
produced, encoding first a fusion between the endogenous protein and -geo
that localizes to the neuronal cell body and second the PLAP protein, which
localizes to the entire cell surface, including the axon.
Left Lower panel: (c) Modified secretory trap insertions into genes encoding proteins with hydrophobic leaders apparently result in fusion proteins that are inserted into membranes in a type I orientation (right) [12]. This places the –galactosidase domain of -geo (blue) in the cytosol, where it is more active than in fusions with leaderless proteins; the latter result in a type II orientation (left) with -galactosidase in the lumen of the endoplasmic reticulum (ER). Right Lower panel: Expression of –geo (blue) and PLAP (purple) in a neuron. |