A suite of Gateway cloning vectors for high-throughput genetic analysis in Saccharomyces cerevisiae.
AuthorsAlberti S, Gitler AD, Lindquist S,
JournalYeast
PubMed ID17583893
'In the post-genomic era, academic and biotechnological research is increasingly shifting its attention from single proteins to the analysis of complex protein networks. This change in experimental design requires the use of simple and experimentally tractable organisms, such as the unicellular eukaryote Saccharomyces cerevisiae, and a range of new high-throughput ... More
Development of R4 gateway binary vectors (R4pGWB) enabling high-throughput promoter swapping for plant research.
We developed a new series of Gateway binary vectors, R4pGWBs, that are plant transformation vectors designed for one-step construction of chimeric genes between any promoter and any cDNA. The structure of R4pGWBs is almost the same as the promoterless type of improved pGWBs (ImpGWBs), except that the attR1 site is ... More
Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells.
AuthorsPaddison PJ, Caudy AA, Bernstein E, Hannon GJ, Conklin DS,
JournalGenes Dev
PubMed ID11959843
RNA interference (RNAi) was first recognized in Caenorhabditis elegans as a biological response to exogenous double-stranded RNA (dsRNA), which induces sequence-specific gene silencing. RNAi represents a conserved regulatory motif, which is present in a wide range of eukaryotic organisms. Recently, we and others have shown that endogenously encoded triggers of ... More
Isolation of rat dihydrofolate reductase gene and characterization of recombinant enzyme.
AuthorsWang Y, Bruenn JA, Queener SF, Cody V,
JournalAntimicrob Agents Chemother
PubMed ID11502523
While assays of many antifolate inhibitors for dihydrofolate reductase (DHFR) have been performed using rat DHFR as a target, neither the sequence nor the structure of rat DHFR is known. Here, we report the isolation of the rat DHFR gene through screening of a rat liver cDNA library. The rat ... More
High throughput immuno-screening of cDNA expression libraries produced by in vitro recombination; exploring the Plasmodium falciparum proteome.
AuthorsKordai Sowa MP, Sharling L, Humphreys G, Cavanagh DR, Gregory WF, Fenn K, Creasey AM, Arnot DE,
JournalMol Biochem Parasitol
PubMed ID14698438
Improved Plasmodium falciparum cDNA expression libraries were constructed by combining mRNA oligo-capping with in vitro recombination and directional cloning of cDNA inserts into a plasmid vector that expresses sequences as thioredoxin fusion proteins. A novel procedure has also been developed for the rapid identification of seropositive clones on high-density filters, ... More
Human protein factory for converting the transcriptome into an in vitro-expressed proteome,.
AuthorsGoshima N, Kawamura Y, Fukumoto A, Miura A, Honma R, Satoh R, Wakamatsu A, Yamamoto J, Kimura K, Nishikawa T, Andoh T, Iida Y, Ishikawa K, Ito E, Kagawa N, Kaminaga C, Kanehori K, Kawakami B, Kenmochi K, Kimura R, Kobayashi M, Kuroita T, Kuwayama H, Maruyama Y, Matsuo K, Minami K, Mitsubori M, Mori M, Morishita R, Murase A, Nishikawa A, Nishikawa S, Okamoto T, Sakagami N, Sakamoto Y, Sasaki Y, Seki T, Sono S, Sugiyama A, Sumiya T, Takayama T, Takayama Y, Takeda H, Togashi T, Yahata K, Yamada H,
JournalNat Methods
PubMed ID19054851
Appropriate resources and expression technology necessary for human proteomics on a whole-proteome scale are being developed. We prepared a foundation for simple and efficient production of human proteins using the versatile Gateway vector system. We generated 33,275 human Gateway entry clones for protein synthesis, developed mRNA expression protocols for them ... More
DNA cloning using in vitro site-specific recombination.
Authors Hartley J L; Temple G F; Brasch M A;
JournalGenome Res
PubMed ID11076863
As a result of numerous genome sequencing projects, large numbers of candidate open reading frames are being identified, many of which have no known function. Analysis of these genes typically involves the transfer of DNA segments into a variety of vector backgrounds for protein expression and functional analysis. We describe ... More