CHE 211 Study Guide - Final Guide: Agarose Gel Electrophoresis, Complementary Dna, Agarose
Organic Chemistry 2: Molecular Genetics
Recombinant DNA
► Over the past 20 years, new methods and techniques called genetic engineering
permitted scientists to cut, splice DNA from different types of cells
► The new synthetic form of DNA, which contains a DNA fragment from another organism
is known as Recombinant DNA this type of technology is used today to form insulin,
antiviral substance called interveron, blood clotting factors (V111) and human growth
hormones
► Most of the work done in Recombinant DNA is done with using E. coli; the DNA present
in bacterial cells have several small circular molecules called plasmids these plasmids,
which are easy to isolate, are capable of replication
► Ability to insert foreign DNA into cells developed after scientists discovered restriction
enzymes
► Restriction enzymes are bacterial enzymes (recognizes specific nucleotide sequences on
a DNA molecule) that cut the sugar-phosphate backbone of DNA at specific nucleotide
sequences the resulting DNA fragment has a sticky end that can form a
phosphodiester bond when they are mixed with the plasmid that are also cut using
restriction enzymes
► Donor (foreign DNA fragment) and plasmid (bacteria) DNA are cleaved by the same
restriction enzyme
► Donor and plasmid DNA are mixed and donor fragment joins to a complimentary
plasmid fragment due to hydrogen bonding
► Plasmid ring is restored using DNA ligase
► Engineered plasmid (recombinant DNA) is introduced to a bacterium to be reproduced
After the foreign DNA fragment joins the plasmid DNA, the recombinant DNA in
the plasmid is able to synthesized the proteins coded by the new DNA
In one day, 1 E.coli bacterium is capable producing a million copies of it,
including the foreign DNA this is known as gene cloning
Agarose Gel Electrophoresis
To study DNA fragment produced by restriction enzyme digestion use agarose gel
electrophoresis
o Using gel electrophoresis, the sizes of each DNA fragment be determined when
compared with the migration pattern of DNA fragments with known sizes
o Digested DNA sample placed in sample slot and electric current is applied
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Document Summary
Over the past 20 years, new methods and techniques called genetic engineering permitted scientists to cut, splice dna from different types of cells. Ability to insert foreign dna into cells developed after scientists discovered restriction enzymes. Donor (foreign dna fragment) and plasmid (bacteria) dna are cleaved by the same restriction enzyme. Donor and plasmid dna are mixed and donor fragment joins to a complimentary plasmid fragment due to hydrogen bonding. Plasmid ring is restored using dna ligase. Engineered plasmid (recombinant dna) is introduced to a bacterium to be reproduced. After the foreign dna fragment joins the plasmid dna, the recombinant dna in the plasmid is able to synthesized the proteins coded by the new dna. In one day, 1 e. coli bacterium is capable producing a million copies of it, including the foreign dna this is known as gene cloning. Southern blotting hybridizes dna fragments already separated on an agarose gel.
