This can pose problems,
as the AMA1-based plasmid has a higher chance of being lost in
progeny than an integrative plasmid. This instability is not constant
in all fungal species that have been studied using these
autonomously replicating plasmids. For instance, mitotic stability
of AMA1-based plasmids is higher in P. nalgiovense than in P. chrysogenum
(Fierro et al., 1996, 2004). In our lab, we have used AMA1-
based plasmids frequently with great success and find that as long
as strains are grown on selective medium, the incidence of plasmid
loss decreases.
As our results demonstrate, using Gateway-enabled vectors
expedites the process of vector construction for genetic techniques.
Not only is Gateway technology widely available and easy to use,
but it also has many advantages over traditional vector construction,
such as increased effectiveness and built-in screening. The
variety of different att sites is ideal for directional cloning of DNA
fragments and can be used to fuse multiple DNA fragments together
or simply attach a tag. We have constructed a series of Gateway-enable
vectors containing various selective markers to streamline genetic
manipulation. These vectors are meant to be used as versatile
genetic tools, as they can be used in a variety of fungal species.
This can pose problems,as the AMA1-based plasmid has a higher chance of being lost inprogeny than an integrative plasmid. This instability is not constantin all fungal species that have been studied using theseautonomously replicating plasmids. For instance, mitotic stabilityof AMA1-based plasmids is higher in P. nalgiovense than in P. chrysogenum(Fierro et al., 1996, 2004). In our lab, we have used AMA1-based plasmids frequently with great success and find that as longas strains are grown on selective medium, the incidence of plasmidloss decreases.As our results demonstrate, using Gateway-enabled vectorsexpedites the process of vector construction for genetic techniques.Not only is Gateway technology widely available and easy to use,but it also has many advantages over traditional vector construction,such as increased effectiveness and built-in screening. Thevariety of different att sites is ideal for directional cloning of DNAfragments and can be used to fuse multiple DNA fragments togetheror simply attach a tag. We have constructed a series of Gateway-enablevectors containing various selective markers to streamline geneticmanipulation. These vectors are meant to be used as versatilegenetic tools, as they can be used in a variety of fungal species.
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