Transformation Lab

Transformation

DNA is released into the environment from a lysed cell and the recipient bacteria cell takes it up acquires observable new trait

Competent

Bacteria capable of taking up DNA

Horizontal gene transfer

Genes passed from an organism to another w/I the same generation

Ice cold calcium chloride

Chemical used by scientists in a laboratory to induce competence of bacterial cells

Plasmid

Small circular double stranded DNA molecule that carries accessory genes separate from those of a bacterial chromosome

Recombinant plasmids

Plasmids that carry additional pieces of DNA that have been inserted through genetic engineering

Has gene of interest

Nonrecombinant plasmid

Does not have gene of interest

Vectors

Plasmids that have been specifically modified and developed for lab purposes b/c they carry foreign DNA into bacteria cells

Antibiotic resistance gene

Allows us to select for bacteria that were successfully transformed

pUC18

Plasmid vector that is used for cloning pieces of foreign DNA in bacteria

2 reasons transformation is difficult

1. It is difficult to get bacteria to take up the plasmid

2. It is difficult to make the plasmid w/ your gene of interest inserted

bla gene

Encodes beta-lactase enzyme (penicillinase)

Enzyme breaks beta lactam ring of penicillin’s

Antibiotic resistance gene (ampicillin)

If bacteria has plasmid they will survive in presence of ampicillin

Product → gives the bacteria w/ plasmid resistant to the antibiotic ampicillin

LacZ gene

Encodes beta galactosidase enzyme

Enzyme breaks down X-gal to form a blue product

Only works if not interrupted by gene of interest

Gene of interest in MCS = no blue

No gene of interest in MCS = blue

Multiple cloning site (MCS)

Insert DNA of interest here

Example: insulin gene would be inserted here