Taq PolymeraseBy: Rachel and Justin

General Protein Synthesis

http://gleesonbiology.pbworks.com/w/page/7537902/M4

1. RNA is copied from DNA through transcription – a process by which the double helix strand is unwound, then copied onto a strand of RNA

http://hyperphysics.phy-astr.gsu.edu/hbase/organic/transcription.html

1. That mRNA then travels outside of the nucleus and into a ribosome

2. The ribosome is like a translator to translate the mRNA into a polypeptide chain.

3. tRNA with an amino acid on one end, and an anticodon on the other, enters the ribosome, and matches with the mRNA, copied from the DNA.

4. As the anticodons and codons match up, the amino acids break away in a polypeptide chain.

http://www.biologycorner.com/bio2/genetics/notes_transcription_translation.html

Enzymes catalyze specific reactions. Each one has an active site that perfectly fits the substrate that the enzyme catalyzes the reaction for. Basically Form Fits Function

Enzymes

http://www.buzzle.com/articles/how-do-enzymes-work.html

Taq Polymerase is a protein composed of a single polypeptide chain consisting of 832 amino acids, taken from the bacterium Thermus Aquaticus, which is thermophilic, meaning it its optimal temperature is extremely high for living organisms; about 75-80°.

Taq Polymerase

http://en.wikipedia.org/wiki/Taq_polymerasehttp://en.wikipedia.org/wiki/File:Thermus_aquaticus.JPG

Bacterium Thermus Aquaticus

It has three domains; a 5′ to 3′ nuclease domain

An inactive 3′ to 5′ exonuclease domain

And a 5′ to 3′ polymerase domain

The inactive exonuclease domain and the polymerase domain together are called the Klentaq fragment.

Structure

•http://www.ncbi.nlm.nih.gov/pubmed/8717047•http://sthor.myweb.uga.edu/bcmb8010/report.pdf

The “palm” section catalyzes the phosphoryl transfer reaction, the transfer of the phosphoryl group (PO3) from a phosphate ester or anhydride to a nucleophile

The “fingers” interact with the incoming nucleoside triphosphate and the template it is being matched to

The “thumb” helps to position the oncoming DNA and enzyme in order to properly carry out the function

The “wrist” section is basically the nuclease domain

Metabolic/Physiological Function

•http://sthor.myweb.uga.edu/bcmb8010/report.pdf•http://www.els.net/WileyCDA/ElsArticle/refId-a0000608.html

Thermus Aquaticus was unintentionally discovered by Thomas D. Brock when

he was examining mushroom samples from Yellowstone National Park Taq polymerase was

discovered in the 1980’s by Kary Mullis in 1983 when he

was adding DNA polymerase to various strands of DNA which greatly increased the

amount of strands presents.

History

•http://www.genetics.org/content/146/4/1207.full.pdf•http://sthor.myweb.uga.edu/bcmb8010/report.pdf•http://pl.wikipedia.org/wiki/Kary_Mullis

Taq polymerase can withstand temperatures needed to produce the best effects in this machine, so scientists can run many PCR cycles automatically.

PCR involves denaturing, annealing and replication steps, usually repeated 20 to 30 times. Denaturing separates the double-stranded DNA into single strands. In the annealing step, primers bind to the segments of DNA to be copied. Taq polymerase goes to work in the replication step: the polymerase builds each single strand of DNA marked by a primer into a new, double-stranded DNA segment.

Genetic testing Infectious disease applications- PCR is used in slow growing bacteria

to allow more DNA to be tested for research

Industrial Purposes - PCR

http://en.wikipedia.org/wiki/Applications_of_PCR

Genetic testing Infectious disease applications-

PCR is used in slow growing bacteria to allow more DNA to be tested for research

Organ transplantation Tissue typing Genetic fingerprinting in

forensics

PCR Uses

•http://en.wikipedia.org/wiki/Applications_of_PCR•http://www.whitcoulls.co.nz/book/organ-transplants/5730966/

Taq Polymerase catalyzes reactions in the nucleus, pertaining to DNA replication and repairs

Taq polymerase utilizes a DNA polymerase domain to catalyze the addition of dNTP to the 3′ hydroxyl terminus of an RNA primer and a 5′ nuclease domain to cleave the downstream, single-stranded 5′ nucleotide displaced by the growing upstream strand

Catalyzed Reaction

http://www.pnas.org/content/102/49/17646.long

http://www.youtube.com/watch?v=_PRUfJBzu-A

Better explanation here, trust us.

Video

Taq polymerase can function at very high temperatures. Sometimes low temperatures cause this enzyme to denature or not function properly.

Some bacteria is harmful to the function of this enzyme Melanin can also affect it Some types of dye or fabrics also inhibit taq polymerase’s function

Environmental Factors

http://www.nfstc.org/pdi/Subject03/pdi_s03_m01_02.htm

Taq Polymerase is used in DNA replication and repairs. It follows the normal function of all enzymes. Taq Polymerase catalyzes the DNA reactions. It is used industrially by the PCR, or the Polymerase Chain Reaction, which is used for genetic testing, fingerprint testing, organ testing, and infection diagnosing. Overall, this enzyme is very useful and helpful in today’s world.

Conclusion