Table 2 |
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Bacterial Assay for the Deubiquitinating Activity of Echinus |
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Plasmid |
Colony Color |
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|
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pUb-Arg-β-Gal |
Blue |
|
pRB105 (yUbp2) |
White |
|
pRB-ec (SF1) |
White |
|
pRB-ec (SF2) |
White |
|
pUb-Arg-β-Gal pRB105 (yUbp2) |
White |
|
pUb-Arg-β-Gal pRB-ec (SF1) |
Blue |
|
pUb-Arg-β-Gal pRB-ec (SF2) |
Blue |
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|
|
|
Table 2. Echinus ec-SF1 lacks deubiquitinating activity on a model substrate in bacteria. Ubiquitin-Arg-β-Gal has a long half-life, and colonies expressing this protein alone are therefore blue in the presence of X-gal substrate. Expression of S. cerevisiae Ubp2 with Arg-β-Gal results in cleavage of ubiquitin, exposing the N-terminus of Arg-β-Gal, which has a short half-life (white colonies). In contrast, expression of ec-SF1 or ec-SF2 does not result in significant cleave Ub-Arg-β-Gal (blue colony color). |
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Copeland et al. BMC Developmental Biology 2007 7:82 doi:10.1186/1471-213X-7-82 |
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