Important

1. Gene Cloning
Host Cell
Prokaryotic
E. coli
B. subtilis
S. cerevisae
Plant cells
Animal cells
Introduction of cloned DNA
Eukaryotic
Transformation
Transfection
Packaging in vitro
Microinjection
Biolistics
An origin of
replication
Selectable
marker(s)
Single
restriction site
Multiple
cloning
sites
Expression
signals
Inserted into a
host cell
Vector or carrier
molecule
Plasmids Phages
Cosmids
Phagemids
Yeast artificial
chromosomes
Viruses
Circular DNA
molecules
Specially
constructed
λ M13
Insertion
vectors
Replacement
vectors
EMPL4
Charon 40
λgt 10
Charon 16A
requires a
Including both
such as such as
need method for
uses a
which
should have may have
and can be
by
Two main types
which are
and have been
such as
such as
such as
two types

2. Cloning strategies
Generation of DNA
fragments
Joining DNA
fragments to
vector
Genomic DNA
Mechanical
shearing
Restriction digests
cDNA
mRNA
Direct chemical
synthesis
Polymerase chain
reaction
DNA fragments for
cloning
Biomolecular
recombinants
Concatemeric
phage/Insert DNA
Blunt end ligation
Use of linkers
Use of adaptors
Homopolymer tailing
Ligation of cohesive termini
cDNA library
Genomic library
Isolate genes
require
need methods for
from either or by
by by synthesis of
gives
gives
by
to give
or
Join to
vector by
produces
used to