"

From 2012.igem.org

Home	Team	Official Team Profile	Project	Parts Submitted to the Registry	Modeling	Notebook	Safety	Attributions

This page is for Guy's research.

Stable/Genomic encoding

http://partsregistry.org/wiki/index.php?title=Part:BBa_K590023

I am currently viewing Yeast integrative plasmid,(YIp), whhich are yeast vectors that utilize integration into the yeast chromosome for survival and are used when studying a DNA gene. http://en.wikipedia.org/wiki/Yeast_artificial_chromosome

http://www.annualreviews.org/doi/pdf/10.1146/annurev.mi.37.100183.001345 http://www.pnas.org/content/75/4/1929.full.pdf http://nar.oxfordjournals.org/content/29/12/e59.full http://www.blackwellpublishing.com/genecloning/pdfs/chapter7.pdf (7.1.3) https://static.igem.org/mediawiki/2008/4/4e/JHU_0708_paper_Shuttle_Vectors_with_Multiple_Unique_Restriction_Sites.pdf

Vectors derived from the 2mm plasmid are called yeast episomal plasmids (YEps). Some YEps contain the entire 2mm plasmid, others include just the 2mm origin of replication. An example of the latter type is YEp13 (Figure 7.3). YEp13 illustrates several general features of yeast cloning vectors. First, it is a shuttle vector. As well as the 2mm origin of replication and the selectable LEU2 gene, YEp13 also includes the entire pBR322 sequence, and can therefore replicate and be selected for in both yeast and E. coli. There are several lines of reasoning behind the use of shuttle vectors. One is that it may be difficult to recover the recombinant DNA molecule from a transformed yeast colony. This is not such a problem with YEps, which are present in yeast cells primarily as plasmids, but with other yeast vectors, which may integrate into one of the yeast chromosomes (p. 135), purification may be impossible.This is a disadvantage because in many cloning experiments purification of recombinant DNA is essential in order for the correct construct to be identified by, for example, DNA sequencing. The standard procedure when cloning in yeast is therefore to perform the initial cloning experiment with E. coli, and to select recombinants in this organism. Recombinant plasmids can then be purified, characterized, and the correct molecule introduced into yeast (Figure 7.4).