Biomedical Engineering Reference
In-Depth Information
Table 9.1
Properties of the different yeast vectors.
Loss in
non-selective
medium
Transformation
frequency
Copy
no./cell
Vector
Disadvantages
Advantages
YIp
10
2
transformants
per
m
g DNA
1
Much less than
1% per
generation
1 Low transformation
frequency
2 Can only be recovered
from yeast by cutting
chromosomal DNA with
restriction endonuclease
which does not cleave
original vector
containing cloned gene
1 Of all vectors, this kind give most
stable maintenance of cloned genes
2 An integrated YIp plasmid behaves
as an ordinary genetic marker, e.g.
a diploid heterozygous for an
integrated plasmid segregates the
plasmid in a Mendelian fashion
3 Most useful for surrogate genetics
of yeast, e.g. can be used to
introduce deletions, inversions and
transpositions (see Botstein &
Davis 1982)
1 Readily recovered from yeast
2 High copy number
3 High transformation frequency
4 Very useful for complementation
studies
1 Readily recovered from yeast
2 High copy number. Note that the
copy number is usually less than that
of YEp vectors but this may be useful
if cloning gene whose product is
deleterious to the cell if produced in
excess
3 High transformation frequency
4 Very useful for complementation
studies
5 Can integrate into the chromosome
1 Low copy number is useful if
product of cloned gene is deleterious
to cell
2 High transformation frequency
3 Very useful for complementation
studies
4 At meiosis generally shows
Mendelian segregation
1 High-capacity cloning system
permitting DNA molecules greater
than 40kb to be cloned
2 Can amplify large DNA molecules
in a simple genetic background
Can get amplification following
chromosomal integration
YEp
10
3
-10
5
transformants
per
m
g DNA
25-200
1% per
generation
Novel recombinants
generated
in vivo
by
recombination with
endogenous 2
m
m
plasmid
Instability of
transformants
YRp
10
4
transformants
per
m
g DNA
1-20
Much greater
than 1% per
generation but
can get
chromosomal
integration
YCp
10
4
transformants
per
m
g DNA
1-2
Less than 1%
per generation
Low copy number makes
recovery from yeast
more difficult than that
with YEp or YRp vectors
YAC
1-2
Depends on
length: the
longer the YAC
the more
stable it is
Stable, since
integrated into
chromosome
Difficult to map by
standard techniques
Ty
Depends on
vector used to
introduce Ty
into cell
~
20
Needs to be introduced
into cell in another
vector
