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Team:Warsaw/Calendar-Main/18 June 2008
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Attribution analyzer
/
Interactive list of topics
Preparation of constructs with OmpA protein fusions
Michał K.
Isolation
of pACYC177.
Digest
of purified pACYC177 with NdeI and BamHI (Tango 2x buffer).
Gel electrophoresis (
Fig. 2
) and
isolation
of proper band (3200 bp).
Clean-up
of OmpA_alpha and OmpA_omega digest products (
Fig. 1
).
Ligation
of purified DNA (pACYC177 with OmpA_alpha and OmpA_omega DNA fragments).
Transformation
of
E.coli
TOP10
with ligation products.
Plating transformants on LB+tetracycline.
Blue/white and rifampicin test
Michał L., Ewa, Marcin
Counting of blue colonies:
Strain
% of blue colonies
pMPMT5
99%
pMPMT5-AID
93%
pMPMT5-AIDT7
81%
pMPMT5-AID+T7
1%
Results of rifampicin test
Strain
Inducer / repressor
Number of colonies on rifampicin
Top10
none
8
Top10 pMPM-AID
0.1% Arabinose
72
Top10 pMPM-AID
none
4
Top10 pMPM-A+T13
0.1% Arabinose
148
Top10 pMPM-A+T13
none
6
Top10 pMPM-AT6
0.1% Arabinose
21
Top10 pMPM-AT6
none
27
Fig. 1. OmpA_alpha (lane 2) and OmpA_omega (lane 3) digest products.
Fig. 2. pACYC177 digested with NdeI and BamHI (lane 2).
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