Abstract:
:Immunization with naked DNA was used to elicit chicken egg yolk antibodies (IgY). Layer hens were inoculated with plasmid DNA encoding the enhanced green fluorescent protein, the fusion protein of Newcastle disease virus, and VP2 of African horse sickness virus. IgY was extracted from egg yolks by polyethylene glycol precipitation. Specific antibodies were present in the yolks of eggs from hens immunized with each of the three different plasmids. This approach to raising polyclonal antibodies obviates the need to produce and purify large quantities of proteins for immunization and can potentially yield large amounts of diagnostically or therapeutically useful reagents.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Romito M,Viljoen GJ,Du Plessis DHdoi
10.2144/01313dd05subject
Has Abstractpub_date
2001-09-01 00:00:00pages
670, 672, 674-5issue
3eissn
0736-6205issn
1940-9818journal_volume
31pub_type
相关文献
BIOTECHNIQUES文献大全abstract::One of the challenges of performing live-cell imaging in plants is establishing a system for securing the sample during imaging that allows for the rapid addition of treatments. Here we report how a commercially available device called a HybriWell™ can be repurposed to create an imaging chamber suitable for Arabidopsi...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/btn-2018-0044
更新日期:2018-06-01 00:00:00
abstract::A simple, nondestructive PCR-based screening method has been developed for identifying putative transgenic soft white winter wheat (Triticum aestivum L.) carrying the coat protein gene of wheat streak mosaic virus. Removal of the endosperm end of individual seed provided sufficient material for DNA extraction and PCR....
journal_title:BioTechniques
pub_type:
doi:10.2144/02323st09
更新日期:2002-03-01 00:00:00
abstract::We have designed and built a magnetic tweezers device that enables the application of calibrated stresses to soft materials while simultaneously measuring their microscale deformation using confocal microscopy. Unlike previous magnetic tweezers designs, our device is entirely portable, allowing easy use on microscopes...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000113701
更新日期:2011-07-01 00:00:00
abstract::Hematopoietic stem cells (HSCs) express Mdr1a/1b and Bcrp1/Abcg2, which are members of the ATP binding cassette transporter family. Mice lacking both Mdr1-type genes (Mdr1a and Mdr1b) or Bcrp1 had normal hematopoietic development, but it has been unclear whether Mdr1a/1b and Bcrp1 play redundant roles in hematopoiesis...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/03356ss04
更新日期:2003-12-01 00:00:00
abstract::We describe a multipurpose eukaryotic expression vector that incorporates the following features: restriction sites for in-frame insertion of cDNAs of interest between sequences encoding the glutathione-S-transferase (GST) and an oligohistidine element, allowing expression of the corresponding fusion proteins; a phosp...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1995-01-01 00:00:00
abstract::Autoantibodies directed against the 68-kDa (U1) ribonucleoprotein antigen are mainly found in sera of patients with mixed connective tissue disease. The corresponding cDNA was fragmented into four regions coding for the major antigenic epitopes A', B', C' and D'. All the epitopes were subcloned and expressed as fusion...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1991-09-01 00:00:00
abstract::Caspase-3 is a key effector caspase that is activated in both extrinsic and intrinsic pathways of apoptosis. Available fluorescent sensors for caspase-3 activity operate in relatively short wavelength regions and are nonoptimal for multiparameter microscopy and whole-body imaging. In the present work, we developed new...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000114377
更新日期:2016-02-01 00:00:00
abstract::We report a modified method of screening for point mutations using a PCR approach based upon the sensitivity of PCR to the 3' terminus of the primer. This method provides a sensitive screen when using either plasmid DNA or bacterial cell lysates. We have optimized the technique for general use to allow rapid screening...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1992-01-01 00:00:00
abstract::Microarray technologies have made possible comprehensive analyses of nucleic acid sequence and expression. However, the technology to obtain efficiently high-quality RNA and DNA suitable for array analysis from purified populations of neoplastic cells from human tissues has not been well addressed. Microdissection can...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/02324rr06
更新日期:2002-04-01 00:00:00
abstract::Reporter assays that use luciferase are widely employed for monitoring cellular events associated with gene expression. In general, firefly luciferase and Renilla luciferase are used for monitoring single gene expression. However, the expression of more than one gene cannot be monitored simultaneously by this system b...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/05386ST03
更新日期:2005-06-01 00:00:00
abstract::We describe a simple and efficient system for epitope mapping by cloning random gene fragments into a specially designed gIIIp-based phage display vector. DNA encoding the antigen of interest is PCR-amplified and partially digested with DNaseI to generate 50-150-bp-long fragments, which are polished with T4 DNA polyme...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/99272st04
更新日期:1999-08-01 00:00:00
abstract::Integral membrane G protein-coupled receptors (GPCRs) compose the single most prolific class of drug targets, yet significant functional and structural questions remain unanswered for this superfamily. A primary reason for this gap in understanding arises from the difficulty of forming soluble, monodisperse receptor m...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000112169
更新日期:2006-05-01 00:00:00
abstract::Phage display holds a key position in the use of combinatorial library approaches for the purpose of protein engineering and discovery. However, modifying the pIII protein of the phage can severely and negatively influence the infectiousness of the phage particle. This concern is particularly relevant when large pIII ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000112724
更新日期:2008-04-01 00:00:00
abstract::For immunological research on the human cytomegalovirus (HCMV), a virus that combines the broad cell tropism of clinical isolates, efficient replication in cell culture, the complete set of MHC-I modulator genes, and suitability for genetic engineering is desired. Here, we aimed to generate a genetically complete deri...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000114606
更新日期:2017-11-01 00:00:00
abstract::We describe a rapid and sensitive method for the detection of nucleotide sequence variation that can be used for large-scale screening of population markers. Denaturing gradient gel electrophoresis (DGGE) detects sequence variants of amplified fragments by the differences in their melting behavior. DGGE detects most s...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/99275rr02
更新日期:1999-11-01 00:00:00
abstract::In this special section of BioTechniques, we examine the role of rapid molecular technologies in the detection and identification of agents of infectious disease (ID) and biological weapons (BWs). Besides the threat posed by the global proliferation of BW technologies, there are numerous emerging and reemerging ID age...
journal_title:BioTechniques
pub_type: 杂志文章,评审
doi:10.2144/03354ss01
更新日期:2003-10-01 00:00:00
abstract::A simple and rapid method for permanently marking autoradiographs is described. This procedure is based on the phosphorescence of light-activated zinc sulfide and the subsequent exposure of x-ray films by this light emission. A lacquer-based carrier allows the zinc sulfide to remain in suspension and permits permanent...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1991-05-01 00:00:00
abstract::Hereditary hemochromatosis (HHC) represents an autosomal recessive disease in which increased iron absorption causes iron overload and irreversible tissue damage. The recently detected association between two point mutations in the HFE gene on chromosome 6p and HHC has made it possible to screen for the disease before...
journal_title:BioTechniques
pub_type:
doi:10.2144/99266st02
更新日期:1999-06-01 00:00:00
abstract::There are little independent data available about how well single nucleotide polymorphism (SNP) genotyping technologies perform in the typical molecular genetics laboratory. We evaluated the utility and accuracy of a widely used technology, template-directed dye-terminator incorporation with fluorescence-polarization ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/02325rr02
更新日期:2002-05-01 00:00:00
abstract::Display of functional antibody fragments on the surface of filamentous bacteriophages allows fast selection of specific phage antibodies against a variety of target antigens. However, enrichment of single chain variable fragment (scFv)-displaying phages is often hampered by the abundance of bacteriophages lacking anti...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/01302rr04
更新日期:2001-02-01 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
pub_type: 杂志文章
doi:
更新日期:1992-01-01 00:00:00
abstract::A simple spectrophotometric method for measuring DNA in proliferating cells is described. The method is an adaptation of the widely used diphenylamine (DPA) reaction to examine DNA in cells grown in a 96-well plate. This assay was capable of detecting as little as 10 ng DNA and could be used to measure DNA in stored a...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1994-07-01 00:00:00
abstract::Silencing of mammalian gene expression by RNA interference (RNAi) technology can be achieved using small interfering RNA (siRNA) or short hairpin RNA (shRNA). However, the relative effectiveness of these two approaches is not known. It is also not clear whether gene-specific shRNA transcribed from an RNA polymerase II...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/04363RR01
更新日期:2004-03-01 00:00:00
abstract::Recent computational and bioinformatics advances have enabled the efficient creation of novel biocatalysts by reducing amino acid variability at hot spot regions. To further expand the utility of this strategy, we present here a tool called Multi-site Degenerate Codon Analyzer (MDC-Analyzer) for the automated design o...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000114177
更新日期:2014-06-01 00:00:00
abstract::We developed a sensitive fluorescence assay for the quantitation of proteins in solution using the NanoOrange reagent, a merocyanine dye that produces a large increase in fluorescence quantum yield upon interaction with detergent-coated proteins. The NanoOrange assay allowed for the detection of 10 ng/mL to 10 microgr...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/03344pt03
更新日期:2003-04-01 00:00:00
abstract::The microcomputer-based image analysis system IB-1000 (developed by Indiana Biotech, Highland, IN) for two-dimensional electrophoresis gels has been described previously (9). It allows the user to compare protein spots between two profiles and identify those spots that are commonly shared in both profiles. This report...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1989-04-01 00:00:00
abstract::Real-time or quantitative loop-mediated isothermal amplification (qLAMP) is a promising technique for the accurate detection of pathogens in organisms and the environment. Here we present a comparative study of the performance of six fluorescent intercalating dyes-SYTO-9, SYTO-13, SYTO-82, SYBR Green I, SYBR Gold, Eva...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000114432
更新日期:2016-07-01 00:00:00
abstract::Ditag genome scanning (DGS) uses next-generation DNA sequencing to sequence the ends of ditag fragments produced by restriction enzymes. These sequences are compared to known genome sequences to determine their structure. In order to use DGS for large-scale genome structural studies, we have substantially revised the ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000113294
更新日期:2009-11-01 00:00:00
abstract::Haplotypes are useful in population genetics and medicine. Determining haplotypes in the absence of DNA samples from appropriate family members can be difficult and laborious. We have developed a rapid and reproducible method for haplotyping an individual in the absence of relatives. The method utilizes pairs of allel...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1991-04-01 00:00:00
abstract::We describe a rapid, simple and inexpensive method for the isolation of DNA from blood clots suited for use in PCR. Our method is based on the lysing and nuclease-inactivating properties of guanidine thiocyanate together with the nucleic acid-binding properties of silica particles. Isolated DNA can be used for in vitr...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1993-02-01 00:00:00