Bacterial Transformation

bacterial transformation

introducing foreign DNA into bacterial cells

Key steps of bacterial transformation

bacteria are treated with CaCl2, heat shock, recovery with nutrient rich luria broth, and placement on antibiotic and arabinose(sugar) covered plates

plasmid

A small, circular DNA molecule found in bacteria, separate from chromosomal DNA, often used in genetic engineering.

orl

origin of replication, top left of plasmid

bla gene(beta lactamase)

allows bacteria to survive on ambicillan plate through resistance, bottom left of plasmid

GFP gene

gene for green flourecent protein, making the bacteria glow, bottom right of plasmid

araC gene

regulates the gene expression of the GFP gene like a switch, actovated by arabinose, top right of plasmid

Luria Broth

nutrient rich medium for bacterial growth

ampicillin

antibiotic that kills bacteria lacking the necessary resistance(bla)

arabinose

sugar which induces GFP expression, making bacteria glow

LB with -pGLO

growth, no glow, since it is normal bacteria with no antibiotic

LB/AMP with -pGLO

no growth, no glo, as bacteria lacks resistance to amp

LB/AMP with +pGLO

growth, no glow, as bacteria has resistance, but lacks gene expression

LB/AMP/ARA with +pGLO

growth and glow, resistance and gene expression

why are plates incubated upside down

to prevent condensation drops from falling and interfering with colonies

why do scientists give their desired bacteria antibiotic resistance

so all bacteria except the ones that they want die