BISC 130 - Chapter 17: Biotechnology

What is Molecular Cloning used for in Molecular Biology Labs?

To get large quantities of pure Protein.

What protein is specifically mentioned in relation to Alzheimer's Disease?

Tau Protein, which is only found in the human brain.

What is the initial step in the molecular cloning process?

Get a Bacterium to take and hold the Gene of interest.

Why do we need a Plasmid in the cloning process?

To ensure the Bacterium recognizes and receives the Gene of interest.

What are plasmids?

Small optional circular DNA molecules that Bacteria recognizes

What happens every time the Bacterium multiplies in molecular cloning?

It multiplies the Gene of interest along with itself.

What is necessary to isolate the Protein of interest in the cloning process?

Purification is required to isolate it from other Proteins - can be done through various methods

What is a common bacterium used for molecular cloning?

E. Coli, because it's easy to keep alive.

What is the purpose of Polymerase Chain Reaction (PCR)?

To artificially amplify the Gene of interest in order to generate millions of copies for analysis..

What are the starting reagents required for the PCR process?

DNA template, Primers, DNA Polymerase, dNDPs, and Buffer/Cofactors.

What happens in STEP 1 of the PCR process?

Heat is applied to unwind double-stranded DNA.

What is the function of DNA Polymerase in PCR?

It extends the Primers and replicates DNA.

What is the problem with DNA Polymerase in STEP 1 of PCR?

It gets denatured and becomes inactive due to high heat.

How did scientists solve the problem of denaturation of DNA Polymerase in PCR?

They used DNA Polymerase from Thermus Aquaticus, a heat-stable organism.

What is the DNA Pol. that is heat resistant and commonly used now?

Taq polymerase, derived from Thermus aquaticus.

What type of environment does Thermus Aquaticus live in?

Hot environments, such as hot springs.

What cuts the plasmid?

Restriction enzymes

At what point do Restriction Enzymes cut the plasmid?

Inverted Repeat Palindrome Sequences

What are Inverted Repeat Palindrome Sequences

Two sequences of nucleotides that are complementary to each other, and are read the same backwards, allowing restriction enzymes to cut DNA at specific sites.