TEV protease-mediated cleavage in Drosophila as a tool to analyze protein functions in living organisms.

abstract：:Genome sequencing projects are either based on whole genome shotgun (WGS) or on a BAC-by-BAC strategy. Although WGS is in most cases the preferred choice, sometimes the BAC-by-BAC approach may be better because it requires a much simpler assembly process. Furthermore, when the study is limited to specific regions of t...

journal_title：BioTechniques

authors： Todesco S,Campagna D,Levorin F,D'Angelo M,Schiavon R,Valle G,Vezzi A

更新日期：2008-01-01 00:00:00

abstract：:Procedures utilizing Chelex 100 chelating resin have been developed for extracting DNA from forensic-type samples for use with the PCR. The procedures are simple, rapid, involve no organic solvents and do not require multiple tube transfers for most types of samples. The extraction of DNA from semen and very small blo...

journal_title：BioTechniques

authors： Walsh PS,Metzger DA,Higuchi R

更新日期：1991-04-01 00:00:00

abstract：:The two-hybrid system is a genetic method to identify proteins that interact with a specific target protein, which is expressed in yeast as a hybrid with a DNA-binding domain. Use of this method entails transforming yeast, both with this DNA-binding domain hybrid and with a library of activation domain hybrids, follow...

journal_title：BioTechniques

authors： Bartel P,Chien CT,Sternglanz R,Fields S

更新日期：1993-06-01 00:00:00

abstract：:The resolution of complex protein mixtures by discontinuous buffer SDS-PAGE is accomplished by their concentration into thin bands in the stacking gel, followed by their separation during migration through the resolving gel. Recombinant human interferon-inducible protein-10 (IP-10), a 10-kDa C-X-C chemokine with four ...

journal_title：BioTechniques

authors： Crow MK,Karasavvas N,Sarris AH

更新日期：2001-02-01 00:00:00

abstract：:Treatment with 2 mM CuSO4 was used to induce a Drosophila melanogaster metallothionein (Mtn) promoter that had been cloned into a recombinant baculovirus. Careful study revealed that the Mtn promoter functioned as an inducible, if somewhat "leaky" promoter within the context of baculovirus-infected cells. In the proce...

journal_title：BioTechniques

authors： Lanier LM,Storm K,Shafaie A,Volkman LE

更新日期：1997-10-01 00:00:00

abstract：:Cycling probe technology (CPT) is a simple signal amplification method for the detection of specific target DNA sequences. CPT uses a chimeric DNA-RNA-DNA probe that is cut by RNase H when bound to its complementary target sequence. In this study, a hybridization assay was developed to detect biotinylated CPT products...

journal_title：BioTechniques

authors： Warnon S,Zammatteo N,Alexandre I,Hans C,Remacle J

更新日期：2000-06-01 00:00:00

abstract：:A streamlined protocol is described that allows high sensitivity antigen detection by Western blotting in a single day. The choice of membrane blotting matrix, as well as blocking reagents, has been optimized in order to allow rapid development of the blot with chemiluminescent reagents. The entire process, from gel t...

journal_title：BioTechniques

authors： Klapper A,MacKay B,Resh MD

更新日期：1992-05-01 00:00:00

abstract：:A simplified method for producing recombinant baculovirus for expression of foreign genes is described. The method utilizes insect cells infected with the wild-type virus before transfection with the plasmid transfer vector, instead of the standard procedure utilizing cotransfection with a plasmid and viral DNA. Recom...

journal_title：BioTechniques

authors： Goswami BB,Glazer RI

更新日期：1991-05-01 00:00:00

abstract：:We report here an improved method for analyzing protein surface expression utilizing a cold-adapted trypsin. Preservation of activity of the enzyme at 0-4°C permits modification of the protease method of surface analysis to temperatures at which trafficking of mammalian plasmalemmal proteins is blocked. This is an imp...

journal_title：BioTechniques

authors： Ahmad F,Coleman SK,Kaila K,Blaesse P

更新日期：2011-04-01 00:00:00

abstract：:Here, we present a rapid and reproducible procedure based on square-wave pulse electroporation that allows efficient penetration of synthetic oligonucleotides into intact yeast cells. This procedure was successfully used to modify the yeast genome with small amounts of oligonucleotide. ...

journal_title：BioTechniques

authors： Barre FX,Mir LM,Lécluse Y,Harel-Bellan A

更新日期：1998-08-01 00:00:00

abstract：:A direct method to study essential genes is to construct conditional knock-down mutants by replacement of their native promoter by an inducible one. In Mycobacteria, replacement of an essential gene promoter with an anhydrotetracycline inducible one was successfully used but required a multi-step approach. In this wor...

journal_title：BioTechniques

authors： Texier P,Coddeville M,Bordes P,Genevaux P

更新日期：2018-09-01 00:00:00

abstract：:Cysts of free-living protozoa have an impact on the ecology and epidemiology of bacteria because they may act as a transmission vector or shelter the bacteria against hash environmental conditions. Detection and localization of intracystic bacteria and examination of the en- and excystment dynamics is a major challeng...

journal_title：BioTechniques

authors： Lambrecht E,Baré J,Claeys M,Chavatte N,Bert W,Sabbe K,Houf K

更新日期：2015-04-01 00:00:00

abstract：:A procedure for amplification by PCR of reproducible allele markers for amplified fragment length polymorphism (Amp-FLP) analysis is presented. We have prepared markers for the allelic products of the VNTR loci D1S80 (MCT118) and D17S30 (YNZ22) and for the hypervariable VNTR locus close to the 3' end of the apolipopro...

journal_title：BioTechniques

authors： Sajantila A,Puomilahti S,Johnsson V,Ehnholm C

更新日期：1992-01-01 00:00:00

abstract：:The relative spatial distribution of cells in a solid tumor contributes to development of malignancy, yet the details of this process remain poorly understood. To elucidate these mechanisms, the ability to extract and analyze the entire DNA content of individual cells whose precise location in the tumor is known is re...

journal_title：BioTechniques

authors： Guo Y,Yang Y,Zhou J,Czajkowsky DM,Liu B,Shao Z

更新日期：2012-04-01 00:00:00

abstract：:An improved Huffman coding method for information storage in DNA is described. The method entails the utilization of modified unambiguous base assignment that enables efficient coding of characters. A plasmid-based library with efficient and reliable information retrieval and assembly with uniquely designed primers is...

journal_title：BioTechniques

authors： Ailenberg M,Rotstein O

更新日期：2009-09-01 00:00:00

abstract：:Blue/white selection is the standard method for detecting a cloned DNA fragment. In the absence of an insert, uninterrupted expression of the vector-encoded alpha-complement of beta-galactosidase (beta-gal), results in the hydrolysis of X-gal (5-bromo-4-chloro-3-indolyl-beta-D-galactoside) and the subsequent blue stai...

journal_title：BioTechniques

authors： Heuermann K,Cosgrove J

更新日期：2001-05-01 00:00:00

abstract：:This report describes a common method of obtaining template DNA from phagemids, phages and plasmids. The strategy is based on the use of the cationic detergent cetyl-trimethylammonium bromide (CTAB) for DNA precipitation. By avoiding phase separation, many manipulation steps are reduced. A time-saving modification to ...

journal_title：BioTechniques

authors： Del Sal G,Manfioletti G,Schneider C

更新日期：1989-05-01 00:00:00

abstract：:A consensus peptide sequence, QSYP, appears as an artifact during the mapping of monoclonal antibodies (MAbs) using a random peptide phage display library. Phage bearing this QSYP sequence were independently selected by four different laboratories screening separate MAb preparations with the same phage library. In eac...

journal_title：BioTechniques

authors： Jacobs JM,Bailey BW,Burritt JB,Morrison SG,Morrison RP,Dratz EA,Jesaitis AJ,Teintze M

更新日期：2003-01-01 00:00:00

abstract：:Mutations in the human MYH7 gene, encoding a slow skeletal muscle/β-cardiac myosin heavy chain, cause different types of myopathies. The nematode model Caenorhabditis elegans has frequently been employed to study the molecular and physiological consequences of MYH7 mutations in muscle function by introducing mutations...

journal_title：BioTechniques

authors： Gil-Gálvez A,Carbonell-Corvillo P,Paradas C,Miranda-Vizuete A

更新日期：2020-06-01 00:00:00

abstract：:Here we present a procedure for preparing amino acid mixtures--having both the desired composition and a physiological pH--at high concentrations for cell-free expression systems. Up to 2.1 mg/mL of active protein was synthesized in batch mode reactions with an all Escherichia coli cell-free expression system. Our met...

journal_title：BioTechniques

authors： Caschera F,Noireaux V

更新日期：2015-01-01 00:00:00

abstract：:Genetic immunization is an attractive approach to generate antibodies because native proteins are expressed in vivo with normal posttranscriptional modifications, avoiding time-consuming and costly antigen isolation or synthesis. Hydrodynamic tail or limb vein delivery of naked plasmid DNA expression vectors was used ...

journal_title：BioTechniques

authors： Bates MK,Zhang G,Sebestyén MG,Neal ZC,Wolff JA,Herweijer H

更新日期：2006-02-01 00:00:00

abstract：:DNA-binding proteins can be purified by their affinity for probes with specific sequences that are tethered to magnetic beads. A restriction site at the end of the tether allows release of probe with factor still on it. This probe with bound factor can be bandshifted directly to compare to bandshifts in whole extracts...

journal_title：BioTechniques

authors： Ren L,Chen H,Sternberg EA

更新日期：1994-05-01 00:00:00

abstract：:A technique that can be used to isolate vector/insert junctions from clones in vectors, such as yeast artificial chromosomes and P1s, and to sequence plasmid inserts more rapidly has been developed. A vector primer is combined with single, randomly chosen oligonucleotides in PCRs, to create pools of products. With 12-...

journal_title：BioTechniques

authors： Swensen J

更新日期：1996-03-01 00:00:00

abstract：:The PCR method has proved to be an invaluable tool for the specific and sensitive detection of genetically modified material (e.g., Roundup Ready Soybean and Bt-176 "Maximizer" Maize) in foodstuffs. The first step in the procedure, namely the purification of nucleic acids from the sample, is often the deciding factor ...

journal_title：BioTechniques

authors： Tengel C,Schüssler P,Setzke E,Balles J,Sprenger-Haussels M

更新日期：2001-08-01 00:00:00

abstract：:We have developed a rapid, sensitive, and quantitative 96-well microplate-based nonradioactive immunoprecipitation/kinase assay to evaluate mitogen-activated protein kinase (MAPK) activity. Three quantitative nonradioactive imunoprecipitation/kinase assays of MAPK were demonstrated on a 96-well microplate coated with ...

journal_title：BioTechniques

authors： Zhang L,Uder S,Juehne T,Brizzard B,Song K

更新日期：2002-02-01 00:00:00

abstract：:Presepsin is a 13-kDa N-terminal glycoprotein of CD14. Previously, agitation-induced increases in presepsin levels have been reported; however, the mechanism remains poorly understood. In this study, we aimed to reveal the mechanism of presepsin increase. The agitated plasma or serum was separated using gel exclusion ...

journal_title：BioTechniques

authors： Morino G,Takahashi G,Kan S,Inoue Y,Sato K,Shirakawa K

更新日期：2021-01-29 00:00:00

abstract：:The systematic preparation of synthetic peptide combinatorial libraries (SPCLs), each composed of tens of millions of peptides that can be screened in existing diagnostically or pharmacologically relevant in vitro assay systems, is reviewed. The identification of optimal peptide sequences has been achieved through the...

journal_title：BioTechniques

authors： Houghten RA,Appel JR,Blondelle SE,Cuervo JH,Dooley CT,Pinilla C

更新日期：1992-09-01 00:00:00

abstract：:Primary human fibroblasts and a series of cell lines (A549, BNL CL.2, H225, NIH 3T3 and Rat-1) are efficiently transfected by using positively charged complexes of plasmid DNA and transferrin-polylysine or polylysine in the presence of glycerol (1 molar to 1.8 molar, depending on the cell type). An increase in gene ex...

journal_title：BioTechniques

authors： Zauner W,Kichler A,Schmidt W,Sinski A,Wagner E

更新日期：1996-05-01 00:00:00

abstract：:In this communication we describe the sequential use of standard and low-melting agarose in a single gel slab for the electrophoresis of DNA. This method has the advantages of high resolution and reproducibility characteristic of standard agarose and the ease of manipulation of DNA for direct cloning, sequential diges...

journal_title：BioTechniques

authors： Fotadar U,Shapiro LE,Surks MI

更新日期：1991-02-01 00:00:00

abstract：:The use of affinity-based tools has become invaluable as a platform for basic research and in the development of drugs and diagnostics. Applications include affinity chromatography and affinity tag fusions for efficient purification of proteins as well as methods to probe the protein network interactions on a whole-pr...

journal_title：BioTechniques

authors： Uhlén M

更新日期：2008-04-01 00:00:00