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Introduction to Recombinant Genetics- Biology 350Index • Syllabus • CourseInfo LogIn • References • Assignment • Next
CLONING VECTORS FOR EUKARYOTES
Vectors For Yeast
The 2 µm plasmid is a naturally occuring 6 kb episomal plasmid found in yeast.
Selection is based on nutrition instead of drugs. Yeast cells that are auxotrophic
(unable to synthesize an essential component, like an amino acid) can only grow when
nutritionally supplemented, or when the missing nutritional function is provided for by
genes contained on a plasmid or extra chromosome.
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
1 of 14 6/16/2011 4:17 AM
Many eukaryotic vectors contain both eukaryotic and prokaryotic signals and can
be replicated in both types of cells. These are called shuttle vectors.
Often, much of the cloning is done in prokaryotic systems and then the recombinant
vectors are grown in eukaryotic cells.
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
2 of 14 6/16/2011 4:17 AM
Why not just express all genes in prokaryotes? Why transfer to eukaryotes for
expression?
YEp Yeast episomal plasmid
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
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YIp - Yeast integrative plasmid
YRps - Yeast replicative plasmid
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
4 of 14 6/16/2011 4:17 AM
YACs - Yeast artificial chromosome (600-1400 kb capacity)
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
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CLONING VECTORS FOR HIGHER PLANTS
Agrobacterium Tumefaciens
The system was derived from the bacteria that causes crown gall disease in plants.
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
6 of 14 6/16/2011 4:17 AM
During infection, the Ti plasmid is integrated into the plant chromosomal DNA.
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
7 of 14 6/16/2011 4:17 AM
The functions of the Ti plasmid have been divided between two plasmid-like vectors
which are co transformed and operate as a binary vector system.
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
8 of 14 6/16/2011 4:17 AM
Transformed plants can be recovered.
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
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A binary vector
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
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Direct gene transfer randomly integrates supercoiled DNA into cells (no homology
required).
CLONING VECTORS FOR ANIMALS
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
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Cloning Into Insect Cells
Drosophilia cells and the P vector
Baculovirus
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
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Cloning In Mammals
Purposes for cloning into animals
Gene knockout
Production of recombinant proteins
Gene therapy
Vectors are often derived from viral components
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
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© 2005 by CA Rinehart Index • Syllabus • CourseInfo LogIn • References • Assignment • Next
This material is intended for use only by WKU students registered for Biology 350. Other uses prohibited.
Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html
14 of 14 6/16/2011 4:17 AM