Shuttle plasmids

Yeast-bacteria shuttle plasmids are usually able to be maintained in both E. coli and yeast and can be episomal or integrated into the host genome. First, plasmid DNA is usually amplified in E. coli before yeast gets transformed. Different antibiotic resistance cassettes are available, and also an abundance of autotrophic markers was established. 

Subcloning into Yeast Expression Vector. Mutant chicken lysozyme sequences located in a 2.5-kilobase (kb) BamHl fragment are subcloned into the Bam HI site of the yeast shuttle plasmid pAB2413 according to the procedure of Gibco BRL Life Technologies, Inc., Grand Island, NY. This... 

We chose first to clone the Xhol restriction fragments containing the XYL gene cassette from pKRD-1,-2,-3,-4 into the Sail site of the 2p-based pUCKmlO yeast -E. coli shuttle plasmid, resulting in the construction of pLNH-31,-32,-33, and -34 (Fig. 3) [15]. These plasmids were collectively designated as the pLNH... 

Fig. 3. The yeast-R. coli shuttle plasmids containing the three xylose-metabolizing gene cassette, KK-AR (or A R)-KD) [15]...

S.T., and Minton, N.P (2009) A mod-uiar system for Clostridium shuttle plasmids. J. Microbiol Methods, 78,... 

In the above, the pSK265 plasmid replicates in both E. coli and a quite different bacterium, B. subtilis. Such plasmids are often referred to as shuttle plasmids. They offer the ability to amplify DNA through propagation in one cell type, and then alter the genotype of another type of organism. 

Denis-Larose, C. Bergeron, H. Labbe, D., et al., Characterization of the Basic Replicon of Rhodococcus Plasmid pSOX and Development of a Rhodococcus Escherichia Coli Shuttle Vector. Applied and Environmental Microbiology, 1998. 64(11) pp. 4363-4367. 

Rousset M, Casalot L, Rapp-Giles BJ, et al. 1998. New shuttle vectors for the introduction of cloned DNA in Desulfovibrio. Plasmid 39 114-22. 

YEP24 Plasmid E. coli, yeast Ampicillin resistance. Yeast shuttle vector... 

There exist a variety of vectors for cloning into eukaryotic systems, ranging from yeast (Saccharomyces as well as Pichia) through insect cells (Baculovims) and plants (Ti plasmid from Agrobacterium tumefaciens) to mammalian cells (transfected by viral or mammalian vectors). As expression in eukaryotic hosts is less efficient than bacterial expression in terms of yield and time and more complicated in terms of vector structure and culture conditions, such eukaryotic expression systems are only used for genes whose proteins require posttranslational modification which is not possible in bacteria. Yeast is the preferred option as a relatively easily culturable single-cell system but posttranslational modification capabilities is limited. The additional complexity can be circumvented in part by exploiting the ability of eukaryotic vectors to act as shuttle vectors, which can be shuttled between two evolutionarily different hosts. Thus, eukaryotic vectors can be replicated and analyzed in bacteria and transfected into eukaryotic cells for expression of the recombinant product. 

Retrieve nucleotide sequences (in fasta format) and restriction maps for one each of bacterial plasmid, cosmid and shuttle vector. 

Fig. 4 Plasmids for the intracellular production of tagged proteins in B. megaterium. All plasmids of the series 1622 are based on the shuttle vector pSTOP1522 (Fig. 3). (a) DNA sequence of ribosome binding site (RBS) and multiple cloning site (MCS) of the expression plasmid pSTOP1622. The...

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