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Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol. Syst. Biol 2:2006.0008

by Tomoya Baba, Takeshi Ara, Miki Hasegawa, Yuki Takai, Yoshiko Okumura, Miki Baba, Kirill A Datsenko, Masaru Tomita, Barry L Wanner, Hirotada Mori , 2006
"... We have systematically made a set of precisely defined, single-gene deletions of all nonessential genes in Escherichia coli K-12. Open-reading frame coding regions were replaced with a kanamycin cassette flanked by FLP recognition target sites by using a one-step method for inactivation of chromosom ..."
Abstract - Cited by 714 (7 self) - Add to MetaCart
We have systematically made a set of precisely defined, single-gene deletions of all nonessential genes in Escherichia coli K-12. Open-reading frame coding regions were replaced with a kanamycin cassette flanked by FLP recognition target sites by using a one-step method for inactivation

Principles of microRNA-target recognition

by Julius Brennecke, Er Stark, Robert B. Russell, Stephen M. Cohen - PLoS Biol , 2005
"... MicroRNAs (miRNAs) are short non-coding RNAs that regulate gene expression in plants and animals. Although their biological importance has become clear, how they recognize and regulate target genes remains less well understood. Here, we systematically evaluate the minimal requirements for functional ..."
Abstract - Cited by 252 (8 self) - Add to MetaCart
for functional miRNA–target duplexes in vivo and distinguish classes of target sites with different functional properties. Target sites can be grouped into two broad categories. 59 dominant sites have sufficient complementarity to the miRNA 59 end to function with little or no support from pairing to the mi

Asymmetry in Flp-mediated cleavage

by Karen H. Luetke, Bao-ping Zhao, Paul D. Sadowski , 1997
"... Flp is a member of the integrase family of site-specific recombinases. Members of the integrase family mediate DNA strand cleavage via a transesterification reaction involving an active site tyrosine residue. The first step of the reaction results in covalent linkage of the protein to the 3′-phospho ..."
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′-phosphoryl DNA terminus, leaving a 5′-hydroxyl group at the site of the nick. We have used Flp recognition target (FRT) sites containing a 5′-bridging phosphorothioate linkage at the site of Flp cleavage to accumulate intermediates in which Flp is covalently bound at a cleavage site. We have probed

Tagging genes with cassette-exchange sites. Nucleic Acids Res 33: e44

by Gilda Cobellis, Giancarlo Nicolaus, Mariangela Iovino, Antonio Romito, Emanuele Marra, Manlio Barbarisi, Marco Sardiello, Francesco P. Di Giorgio, Nicola Iovino, Massimo Zollo, Andrea Ballabio, Riccardo Cortese , 2005
"... In an effort to make transgenesis more flexible and reproducible, we developed a system based on novel 50 and 30 ‘gene trap ’ vectors containing hetero-specific Flp recognition target sites and the corres-ponding ‘exchange ’ vectors allowing the insertion of any DNA sequence of interest into the tra ..."
Abstract - Cited by 8 (1 self) - Add to MetaCart
In an effort to make transgenesis more flexible and reproducible, we developed a system based on novel 50 and 30 ‘gene trap ’ vectors containing hetero-specific Flp recognition target sites and the corres-ponding ‘exchange ’ vectors allowing the insertion of any DNA sequence of interest

Combinatorial approaches to finding subtle signals in DNA sequences,”

by Pavel A Pevzner , Sing-Hoi Sze - in Proceedings of the 8th International Conference on Intelligient Systems for Molecular Biology, , 2000
"... Abstract Signal finding (pattern discovery in unaligned DNA sequences) is a fundamental problem in both computer science and molecular biology with important applications in locating regulatory sites and drug target identification. Despite many studies, this problem is far from being resolved: most ..."
Abstract - Cited by 258 (5 self) - Add to MetaCart
Abstract Signal finding (pattern discovery in unaligned DNA sequences) is a fundamental problem in both computer science and molecular biology with important applications in locating regulatory sites and drug target identification. Despite many studies, this problem is far from being resolved

FLP-mediated DNA mobilization to specific target sites in Drosophila chromosomes

by Robert B Petersen, See Profile, Mary M. Golic, Yikang S. Rong, Robert B. Petersen, S. L. Lindquist, Kent G. Golic , 1997
"... FLP-mediated DNA mobilization to specific target sites in Drosophila chromosomes ..."
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FLP-mediated DNA mobilization to specific target sites in Drosophila chromosomes

Chimeras of the Flp and Cre recombinases: tests of the mode of cleavage by Flp and Cre

by A C Shaikh , Paul D Sadowski - J Mol Biol
"... The Flp and Cre recombinases are members of the integrase family of tyrosine recombinases. Each protein consists of a 13 kDa NH 2 -terminal domain and a larger COOH-terminal domain that contains the active site of the enzyme. The COOH-terminal domain also contains the major determinants for the bin ..."
Abstract - Cited by 5 (0 self) - Add to MetaCart
speci®cities in that they bind strongly to hybrid sites containing elements from both the Flp and Cre DNA targets but poorly to the native target sites. In this study we have taken advantage of the unique binding speci®ci-ties of Fre and Clp to examine the mode of cleavage by Cre, Flp, Fre and Clp. We

Chemical probe and missing nucleoside analysis of Flp recombinase bound to the recombination target sequence

by Amy Kimball, Melissa L. Kimball, Makkuni Jayaram, Thomas D. Tullius , 1995
"... The Flp protein catalyzes a site-specific recombination reaction between two 47 bp DNA sites without the assistance of any other protein or cofactor. The Flp recognition target (FRT) site consists of three nearly identical sequences, two of which are separated by an 8 bp spacer sequence. In order to ..."
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The Flp protein catalyzes a site-specific recombination reaction between two 47 bp DNA sites without the assistance of any other protein or cofactor. The Flp recognition target (FRT) site consists of three nearly identical sequences, two of which are separated by an 8 bp spacer sequence. In order

Construction of Escherichia coli K-12 Strain Deficient in relA and spoT Using the λ Red Site-Specific Recombinase System

by So Ra Lee
"... Previous studies have shown that an exposure to sub-inhibitory levels of kanamycin induces capsule synthesis which confers antibiotic resistance. To examine the role of spoT and relA genes in capsule synthesis and acquired physiological antibiotic resistance during sub-inhibitory kanamycin treatmen ..."
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resistance gene that is flanked by FLP recognition target sites. This product was electroporated into the strain JW2755-3 expressing the λ Red recombinase system. Five chloramphenicolresistant transformants were selected and PCR-tested for the presence of new junctions and locus-specific fragments. Construct

FLP-mediated DNA mobilization to specific target sites in Drosophila chromosomes. Nucleic Acids Res

by Mary M. Golic, Yikang S. Rong, Robert B. Petersen, S. L. Lindquist, Kent G. Golic , 1997
"... The ability to place a series of gene constructs at a specific site in the genome opens new possibilities for the experimental examination of gene expression and chromosomal position effects. We report that the FLP–FRT site-specific recombination system of the yeast 2µ plasmid can be used to integra ..."
Abstract - Cited by 12 (0 self) - Add to MetaCart
to integrate DNA at a chromosomal FRT target site in Drosophila. The technique we used was to first integrate an FRT-flanked gene by standard P element-mediated trans-formation. FLP was then used to excise the FRT-flanked donor DNA and screen for FLP-mediated re-integration at an FRT target at a different
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