Laboratory: purify plasmid

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Laboratory purify plasmid restriction Lecture
restriction mapping In-Class Writing
rewrite sentences (pages 60-61) Hand In flow
chart 1 Read Appl. Environ. Microbiol. 63
4920-8, 1997
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cut with SalI
5G/TCGA C3 3C AGCT/G5
cut with SalI
cut with SalI
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pBR322
5 TCGA
AGCT 5
Mix SalI restriction fragments. Add DNA ligase
and ATP.
lux operon
5 TCGA
AGCT 5
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Transform ligated DNA into E. coli. Select
ampicillin-resistant colonies. Only circular
molecules survive RecBCD nuclease destroys
linear DNA. Transformants contain vector or
vector lux (either orientation).
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Reporter Gene
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Transgenic tobacco leaves expressing luciferase
Image courtesy of Wayne M. Barnes
veins bright capillaries
veins dark
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wound-inducible transcription factor
wound
plant promoter
luciferase gene
transcription
RNAPol
lux mRNA
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Transgenic tobacco leaves expressing luciferase
Image courtesy of Wayne M. Barnes
Veins Dark
unwounded wounded
Wound Inducible
unwounded/wounded
unwounded/wounded
unwounded
wounded
Capillaries
Veins Bright