Protocol tips
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To test independent induction of two genes, GFPmut2 in pJS102 was replaced by mScarlet-I (amplified from a plasmid [19]) to make pDG101. Plasmids were cotransformed into MG1655 by electroporation according to the above protocol, except with 1 μl (each) undiluted plasmid (20 to 40 ng) and selection on LB agar plates with both spectinomycin and carbenicillin. |
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| Protocol tips |
| To test independent induction of two genes, GFPmut2 in pJS102 was replaced by mScarlet-I (amplified from a plasmid [19]) to make pDG101. Plasmids were cotransformed into MG1655 by electroporation according to the above protocol, except with 1 μl (each) undiluted plasmid (20 to 40 ng) and selection on LB agar plates with both spectinomycin and carbenicillin. |
Publication protocol
To test independent induction of two genes, GFPmut2 in pJS102 was replaced by mScarlet-I (amplified from a plasmid [19]) to make pDG101. Plasmids were cotransformed into MG1655 by electroporation according to the above protocol, except with 1 μl (each) undiluted plasmid (20 to 40 ng) and selection on LB agar plates with both spectinomycin and carbenicillin. Sequence maps are included in an online repository (33), and plasmids useful for constructing additional two-gene expression systems (pJS101 and pJS102) (Table 1) are available from Addgene (deposit no. 118280 and 118281) and have been verified by whole-plasmid sequencing (34).
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