Topic 11 - Biotechnology

DNA technology

techniques used to isolate, purify, analyze, and manipulate DNA sequences; genetic engineering

GMO

genetically modified organism whose genome has been engineered to introduce or change a genetically trained trait

biotechnology

any technique used to modify a biological system; includes genomics and bioinformatics

restriction enzymes

also known as endonucleases; cuts DNA at restriction sites, creating restriction fragments and single stranded sticky ends

single stranded sticky ends aim to..

find its complementary restriction fragment

gene cloning

making multiple copies of a gene and inserting it into a host cell

cloning vectors

plasmid used to insert a DNA molecule for cloning

a GMO is an example of

a recombinant bacterium

DNA wanted to clone and plasmid are cut with the…

same sticky end restriction enzyme

cut DNA and cut plasmid are sealed by

DNA ligase

recombinant plasmid is reintroduced into bacteria through…

transformation

recombinant plasmid

plasmid with the new gene of interest

PCR

polymerase chain reaction; process used to get the gene of interest

PCR steps

denaturing, annealing, extension

denaturing

96 degrees C; double strand to single strand

annealing

55-65 degrees C; primers match to complementary sequences at other end

extension

72 degrees C; target sequences repliacted into 2 copies of original strand

exponential growth

a couple of rounds of PCR giving tens of thousands of copies of desired gene

gel electrophoresis

DNA has negative charge and will move to other side with positive end; smaller fragments move faster and larger fragments move slower

agarose electrophoresis

used to compare PCR results using a standard ladder (control) to compare movement of segments and how far they travelled

ecoli that did not take introduced plasmid with grow..

blue

ecoli that did take introduced plasmid will grow…

clear

cDNA

used for cloning eukaryotic cells; uses reverse transcriptase to make DNA from RNA and is complementary to a mRNA template

central dogna

DNA to RNA to protein

stem cells

type of genetic engineering; undifferentiated precursor cells that can turn into different cell types

totipotent stem cells

can turn into fully functioning organism (very first cell)

pluripotent stem cells

have potential to turn into nearly every cell type (embryonic structure)

multipotent

can turn into handful of cell types (adult cell types)