Shuttle vectors

A variety of tiansformation techniques using E. co/ -ye st shuttle vectors and yeast selectable markets, as well as efficient yeast promoters and signal... 

McLuckie, K. I. Routledge, M. N. Brown, K. Gaskell, M. Farmer, P. B. Roberts, G. C. Martin, E. A. DNA adducts formed from 4-hydroxytamoxifen are more mutagenic than those formed by alpha-acetoxytamoxifen in a shuttle vector target gene replicated in human Ad293 cells. Biochemistry 2002, 41, 8899-8906. 

Another report describing an approach to achieve alleviation of sulfur repression came from the Matsui research group. The dsz genes were cloned into a strain Rhodococcus sp. strain T09 under the promoter rrn of the strain T09 using a Rhodococcus-E. coli shuttle vector [214,215], This resulted in a strain which desulfurized DBT to 2-HBP in presence of sulfate, cysteine, or methionine. Similar approach was also used by Kurane to construct a gene expressing dszA-D enzymes, which eliminate the sulfate inhibition effects [216],... 

Another group from Japan also successfully introduced additional gene copies of dszABC and dszD into the host strain and improved desulfurization activity by 2 1 fold [207], The dsz genes were introduced via a Rhodococcus-E. coli shuttle vector pRHKl into R. erythropolis KA2-5-1 (Table 11). 

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. 

The construction of a shuttle vector (pSM73) which can be replicated both in Rhodococcus and in E. coli containing a gene reporter without its own promoter and on the cloning, before such gene, of random fragments of chromosomal DNA of Rhodococcus consists of ... 

The target gene is markedwith yellow, transposon is blue and the shuttle vector is indicated with a circle. The vector delivering the transposon will be eliminated after the random insertion of the transposable element. Insertion of the transposon inactivates the target gene function this can be corrected via introduction of the gene on a vector (complementation). 

Shrink-resist science/technology development of, 26 391 Shrink-resist treatments, 26 391-393 additive, 26 393 chlorine-based, 26 392 chlorine-free, 26 392-393 Shuiskite, 6 471t Shutdown period, 29 494 Shutdown systems, 20 671-672 Shuttle vectors, 26 482-483 Sialon-bonded silicon carbide, 22 541 Siberian red lead, 6 468 S-iB-S block copolymers, 24 707 SiC-ceramic, 22 525. See also Silicon carbide... 

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. 

Staub, J.M. and Maliga, R. (1995). Marker rescue from the Nicotiana tabacum plastid genome using a plastid Escherichia coli shuttle vector. Mol. Gen. Genet. 249 37 2. 

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

Dees, C.J. Travis, C. (1996) Phenotypic and genotypic analysis of rat liver epithelial cells infected with retroviral shuttle vectors. Cancer Lett., 107, 19-28... 

West, S. E. H., Schweizer, H. P., Dali, C., Sample, A. K. Runyen-Janecky, L. J. (1994). Construction of improved Escherichia—Pseudomonas shuttle vectors derived from pUC18/19 and sequence of the region required for their replication in Pseudomonas aeruginosa. Gene, 128, 81-6. 

Shuttle Vectors Can Be Cloned into Cells of Different Species Constructing a Library ... 

Vectors that include replication systems derived from more than one host species are known as shuttle vectors. Such vectors commonly include a replication system able to function in E. coli and one that works in a second host, which may be bacterial or eukaryotic. Initial cloning and amplification of the DNA segment to be studied is often carried out in E. coli because it is easier to make large quantities when culturing in E. coli. The recombinant DNA molecule, consisting of the bifunctional vector plus the cloned segment of DNA, is then introduced into the second host, where the purpose is usually to measure the expression of the genes carried by the vector. Shuttle vectors that can replicate in both E. coli and yeast are the most common. 

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. 

Figure 9.2. Shuttle vector map retrieved from Riken Gene Bank. The map of shuttle vector pYAC 3/4/5 shows major restriction sites.

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

An expression vector for//, volcanii was constructed by Nieuwlandt and Daniels (1990) by incorporating the promoter of the H. volcanii tRNA gene into a derivative of the H. volcanii shuttle vector pWL102 (Lam and Doolittle, 1989). The applicability of this vector is limited,... 

Lutfalla G, Armbruster L, Dequin S, Bertolotti R (1989), Construction of an EBNA-producing line of well-differentiated human hepatoma cells and of appropriate Epstein-Barr virus-based shuttle vectors, Gene 76 27-39. 

Fig. 4. Schematic representation of yeast one-hybrid transcription factor screening. C. roseus cDNAs were cloned in a fusion with the GAL4-activation domain (GAL4-AD) in yeast/E.coli shuttle vector pACTII. Yeast reporter strains carrying a tetramer of the cis-acting elements of interest (indicated in the figure with 4xbait) were transformed with the cDNA library and selected for transformation and reporter gene activation on medium lacking leucine and histidine, respectively...

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