Analyzing the Structure and Function of Genes

Chapter 10 - Essential Cell Biology 6th Edition by Bruce Alberts, Rebecca Heald, Karen Hopkin, Alexander Johnson, The Estate of Julian Lewis, David Morgan, Martin Raff, Keith Roberts, and Peter Walter

Biotechnology

The science of using living systems to benefit humankind

Genetic engineering

Direct alteration of an organism’s genetics

Recombinant DNA

Genetic engineering is the result of introducing new DNA into a host

transgenic

If the DNA introduced comes from a different species, the host organism is called …

Insulin

• ______ introduced into a bacterium via:

  • Mating or conjugation

  • Viral infection or transduction

  • Taking up DNA from environment or transformation

recognition sites

Molecular cloning relies on specific regions or sequences of DNA

restriction endonucleases/enzymes

Recognition sites are detected and altered by what enzymes?

• they protect bacteria from foreign bacteria

E. coli

In Hershey and Chase experiments:

Bacteriophage T2 kills _____ and only has DNA and protein in its structure

palindromic

RE recognizes _______ sequences of DNA.

sticky ends

they have complimentary over hangs

blunt ends

no over hangs

PCR (polymerase chain reaction)

Scientists use ______ to create the restriction sites near the start and stop codon.

• •Gene of interest is excised or “cut” using restriction enzymes

Vectors (Plasmid)

used to deliver into a host

Plasmids

______ must contain:

• Multiple cloning site (MCS)

• Reporter gene

• Selective marker → antibiotic marker amp

insert

A short sequence containing multiple unique restriction enzyme recognition sites that are used to ____ DNA into the plasmid.

Reporter gene

• artificially engineered into the plasmid

• encodes a protein that allows for visualization of DNA insertion.

lacZ

encodes for beta-galactosidase

beta-galactosidase

breaks down lactose

radioactive isotopes; fluorescent dyes

DNA probes are tagged with molecular markers, such as ________ or __________, to visualize where the probe binds

Southern blotting

used to detect DNA

• Requires a probe that is complementary to the sequence of interest

larger; heavier

Resulting recombinant DNA is ______ and ______ than the vector or insert alone.

Microarray analysis

mRNA from sample is isolated and changed into complimentary DNA (cDNA) via reverse transcriptase

Reverse transcription

process by which complementary DNA (cDNA) is made

• crucial part of the lifecycle of such viruses

• found in viruses that have to convert their RNA genome into DNA

Northern blot

is used to detect RNA