Nonessential region Cosmids: plasmid vectors containing foreign DNA Digestion w ends restrictio plus only the cos(cohesive end) site from the lambda genome Hynd DNA Sizes of a cloning gene the Packaging with vectors can carry YAC>Cosmid>Lambda> Plasmids
Cosmids: plasmid vectors containing foreign DNA plus only the cos (cohesive end) site from the lambda genome Sizes of a cloning gene the vectors can carry: YAC>Cosmid>Lambda>Plasmids
Other vectors Yeast artificial chromosomes (YAC) TEL ARS CEN Not I Not I URA3 INSERTED DNA YAC vectors are only about 10 KB, but can carry 200-800 KB DNA sequences YAC has been developed and used for the human genome project
Other Vectors Yeast artificial chromosomes (YAC) YAC vectors are only about 10 KB, but can carry 200-800 KB DNA sequences YAC has been developed and used for the human genome project
Vectors for DNA Sequencing: Bacteriophage M13 Phagemid: a hybrids ICG AGC TCG GTA CCC GGG GAT CCT CTA GAG TCG ACC TGC AGG CAT GCA A between a Asn SerSerSer Val Pro Gly Asp Pro Leu Glu Ser Thr Cys Arg His Ala Ser filamentous phage, like Mi3 and a plasmid Vectors with cloned DNA Vectors without cloned dNA X-gal: 5-bromo-4 hloro-3-indolyl-B-D galactopyranoside
Vectors for DNA Sequencing: Bacteriophage M13 Phagemid: a hybrids between a filamentous phage, like M13 and a plasmid X-gal: 5-bromo-4- chloro-3-indolyl-b-Dgalactopyranoside Vectors without cloned DNA Vectors with cloned DNA
Other specialized vectors Expression vectors: to obtain synthesis of the protein coded for by the foreign gene cloned into the vector Secretion vectors: protein product is not only expressed but secreted (excreted) from the cell Shuttle vectors: move dna between unrelated organisms, can replicate in two different organisms
Other specialized vectors Expression vectors: to obtain synthesis of the protein coded for by the foreign gene cloned into the vector. Secretion vectors: protein product is not only expressed but secreted (excreted) from the cell. Shuttle vectors: move DNA between unrelated organisms, can replicate in two different organisms
Hosts for cloning vectors Ideal hosts: rapid growth, capable of growth in cheap culture medium, not harmful or pathogenic transformable by dNa, stable Prokaryotic hosts: E coli, Bacillus subtilis Eukaryotic hosts: Saccharomyces cerevisiae DNA vIrus sv40. a virus causing tumors in primates, can be used as a cloning vector into human culture lines Retroviruses. vaccinia virus are useful too Baculovirus. an insect DNA virus can be used to transfer dna to insect cell lines
Hosts for cloning vectors Ideal hosts: rapid growth, capable of growth in cheap culture medium, not harmful or pathogenic, transformable by DNA, stable. Prokaryotic hosts: E. coli, Bacillus subtilis. Eukaryotic hosts: Saccharomyces cerevisiae. DNA virus SV40, a virus causing tumors in primates, can be used as a cloning vector into human culture lines. Retroviruses, vaccinia virus are useful too. Baculovirus, an insect DNA virus can be used to transfer DNA to insect cell lines