MBIO 4440 Lecture 4: Cell Cycle in Caulobacter
20 Jan 2019
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Document Summary
This system allows tracking any site in the chromosome by time-lapse fluorescence microscopy. The system is based on the delivery of the lac operator site laco together with a selection marker (in this case a kanamycin resistance cassette) close to the dna region of interest. Laco is a dna region that tightly binds the transcriptional regulator laci. The strain can be created where the laco site is delivered with the antibiotic marker at a particular location in the chromosome by homologous recombination as shown in figure a. (the. Laco site can also be delivered randomly by transposon mutagenesis; we will study this in unit. Proteins are written in upper case no italics. Next, a plasmid containing the gene encoding the transcriptional regulator laci (fused to a fluorescent protein gene) under the control of an inducible promoter is introduced. In the figure, the cyan fluorescent protein (cfp) is used. The construct and the laci-cfp fusion protein are shown in figure b.
