U - Genetic Engineering

Growth hormone is produced by the ____ and regulates ____ ____

pituitary, body size

1st stage of Engineering hGH-producing bacteria

design and build a plasmid

2nd stage of Engineering hGH-producing bacteria

transform bacteria

3rd stage of Engineering hGH-producing bacteria

screening and selection

expression vector

designed to express a transgene, and has all the features of a cloning vector

Bacterial gene expression machinery are compatible with bacterial regulatory sequences (______, ______ ______ sequences, etc)

promoters, transcription termination

bacteria cannot splice out 

introns

Many bacterial expression vectors have the polylinker/MCS located inside the ____ gene

lacZ

The transcript produced will retain key

features of the lacZ _____ (e.g. a Shine- Dalgarno sequence)

5’ UTR

Insertion of a transgene ______ expression of functional lacZ from the plasmid (enables blue-white screening)

prevents

To isolate our gene of interest, we can do ___ to amplify the specific sequence we want

PCR

Because we want the hGH ORF without any introns, we should use ____ rather than genomic DNA

cDNA

Once we isolate our desired insert by PCR,

we can insert it into the polylinker/MCS region

Selection and screening

strategies geneticists use to identify the bacteria of interest

selection

Only those that satisfy a certain criterion survive

screening

Evaluate to see which that satisfy a certain criterion