Abstract:
:We report a robust method for studying en masse changes in translation using cDNA arrays. The relative distribution of messenger RNAs (mRNAs) along polysome gradients was monitored by performing cDNA array analysis of each gradient fraction and quantifying the mRNA translational status by regression analysis. Using this strategy to study human carcinoma cells exposed to short-wavelength ultraviolet light (UVC), we identified a subset of 17 translationally induced mRNAs and a subset of 69 translationally repressed mRNAs following UVC irradiation. We describe an effective approach for globally investigating changes in protein biosynthesis.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Mazan-Mamczarz K,Kawai T,Martindale JL,Gorospe Mdoi
10.2144/05391ST01subject
Has Abstractpub_date
2005-07-01 00:00:00pages
61-2, 64, 66-7issue
1eissn
0736-6205issn
1940-9818pii
05391ST01journal_volume
39pub_type
杂志文章相关文献
BIOTECHNIQUES文献大全abstract::[Formula: see text] Known to be a sturdy weapon in a scientist's arsenal, how has the gene editing tool CRISPR been applied in the fight against COVID-19? ...
journal_title:BioTechniques
pub_type: 新闻
doi:10.2144/btn-2020-0145
更新日期:2020-11-01 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::Antibiotics are widely useful in medicine, agriculture, and industrial fermentations. However, increasing problems with resistant strains call for restrained use and alternative strategies. Antisense peptide nucleic acids (PNAs) show potent bactericidal effects when targeted against the essential Escherichia coli acpP...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:2003-11-01 00:00:00
abstract::A method for cloning single-stranded oligonucleotides in a plasmid vector has been developed. The method relies on ligation of the oligonucleotide into suitable restriction enzyme sites of the cloning vector such that the site at the 5' end has a 5' overhang [for example, a Bgl II site (A decreases GATCT)], and the si...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1989-04-01 00:00:00
abstract::Currently, in real-time PCR, one often has to choose between using a sequence-specific probe and a nonspecific double-stranded DNA (dsDNA) binding dye for the detection of amplified DNA products. The sequence-specific probe has the advantage that it only detects the targeted product, while the nonspecific dye has the ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000112101
更新日期:2006-03-01 00:00:00
abstract::A brief description of the setup of an inexpensive densitometric analysis system is provided. This system uses a software package and scanner that can be obtained for a total of less than $1000 and that can be operated by any Macintosh computer, including a MacPlus. In the event that a scanner is already available, th...
journal_title:BioTechniques
pub_type:
doi:
更新日期:1994-06-01 00:00:00
abstract::Here we report the construction of a histidine-tagged T4 RNA ligase expression plasmid (pRHT4). The construct, when overexpressed in BL21 (DE3) cells, allows the preparation of large quantities of T4 RNA ligase in high purity using only a single purification column. The histidine affinity tag does not inhibit enzyme f...
journal_title:BioTechniques
pub_type:
doi:10.2144/02336st03
更新日期:2002-12-01 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
pub_type: 杂志文章
doi:10.2144/btn-2020-0136
更新日期:2021-01-29 00:00:00
abstract::We have developed a novel method for the rapid preparation of large quantities of 3' end-labeled single-stranded (ss) DNA (ssDNA) probes for transcript mapping. A recombinant phagemid vector containing the probe sequence was used to raise large quantities of ssDNA. Based on the DNA sequence of the probe, an oligonucle...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1993-05-01 00:00:00
abstract::A new procedure is described for the generation of high-titer, helper-free retrovirus vectors employing receptor-mediated, adenovirus-augmented transfection into a standard packaging cell line. Viral titers are increased 30-fold to 100-fold in transiently (> 10(5) infectious units per mL) and stable (> 10(7) infectiou...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1995-03-01 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
pub_type: 杂志文章
doi:10.2144/000114274
更新日期:2015-04-01 00:00:00
abstract::Random mutagenesis followed by an in vitro selection procedure was shown to be capable of identifying important bases of the hairpin ribozyme for cleavage of an RNA target sequence. The selection scheme enriched the RNA population for those molecules capable of efficient site-specific self-cleavage in the absence of l...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/98242st05
更新日期:1998-02-01 00:00:00
abstract::Previous work from this laboratory has shown that immunoporation has the potential for the selective transfection of a range of different animal cells based on their immunological identity. The unique ability of immunoporation to target cells for transfection combined with the high efficiency of transfection and the h...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/04372RR05
更新日期:2004-08-01 00:00:00
abstract::Large genes present particular cloning difficulties, especially when expressed at relatively low levels. We describe a novel method, termed 3' rapid amplification of cDNA ends (RACE) walking, for the rapid determination of unknown 3' flanking sequence of a large cDNA. The technique is a derivative of the anchored PCR ...
journal_title:BioTechniques
pub_type:
doi:10.2144/03344st04
更新日期:2003-04-01 00:00:00
abstract::Plasmid pUC18rspL is a 3.788-kilobase pair vector, derived from pUC18, pKK232-8 and pHSG664, which identifies promoter-bearing DNA fragments functional in StrA E. coli by activation of a promoterless streptomycin-sensitive gene cartridge (rspL). Expression of the plasmid-borne rspL gene leads to sensitivity dominance ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1990-12-01 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
pub_type: 杂志文章
doi:10.2144/01312pf01
更新日期:2001-08-01 00:00:00
abstract::We have developed a high-throughput, multiplex reverse transcription PCR (RTPCR) assay that is suitable for the analysis of medium-to low-copy cellular RNA transcripts from small numbers of cells (10(4)). High throughput was attained by utilizing microplate-based RNA extraction and RTPCR protocols, followed by PCR pro...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/97226st02
更新日期:1997-06-01 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::Based on the requirement of a glutathione-deficient mutant strain of Saccharomyces cerevisiae to take up external glutathione for growth on synthetic media, a simple agar diffusion test for quantitative detection of total glutathione from various sources was established. Glutathione concentrations can be reliably dete...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/96215st07
更新日期:1996-11-01 00:00:00
abstract::Real-time PCR is becoming a preferred method for quantification of minute amounts of nucleic acids. To achieve the full potential of this technique, accurate and convenient models for post-PCR data analysis are required. In this study, three different models were chosen to quantify the definitive copy numbers of Cucum...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000112750
更新日期:2008-06-01 00:00:00
abstract::Cell-based immunizations are often used when membrane antigens are difficult to purify. To confirm that an antibody binding to the surface of a cell line is, in fact, binding to the desired antigen, FACS can be performed independently on two cell lines, a transfected cell line expressing the antigen of interest and a ...
journal_title:BioTechniques
pub_type:
doi:10.2144/02323dd05
更新日期:2002-03-01 00:00:00
abstract::Chitin from crustacean shells has often been used to isolate and purify plant lectins that have an affinity for poly-N-acetylglucosamine (poly-GlcNAc). When we used washed chitin from crab shells as an affinity medium to isolate a lectin from Pinus strobus L. (eastern white pine) ovules, we found that a substance havi...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1992-02-01 00:00:00
abstract::Expanding applications of cDNA microarrays such as fine needle aspiration biopsy and laser capture microdissection necessitate the ability to perform arrays with minute starting amounts of RNA. While methods for amplifying RNA have been advocated, the fidelity of array results using amplified material has not been ful...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/02334mt04
更新日期:2002-10-01 00:00:00
abstract::New cloning vectors have been developed with features to enhance quick allelic exchange in gram-negative bacteria. The conditionally replicative R6K and transfer origins facilitate conjugation and chromosomal integration into a variety of bacterial species, whereas the sacB gene provides counterselection for allelic e...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/btn-2020-0135
更新日期:2021-01-25 00:00:00
abstract::We previously reported that cervicovaginal lavages (CVL) contain a factor that enhances the replication of human immunodeficiency virus (HIV) by increasing virus transcription in T cells and monocytoid cells. This factor was named the HIV-inducing factor (HIF). To determine the molecular mass of HIF, we adapted a blot...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/00283st05
更新日期:2000-03-01 00:00:00
abstract::We report a straightforward methodology for purification of recombinant proteins by incorporating a short hydrophilic peptide marker segment at their N-termini. A calcium-dependent antibody that reacts primarily with the first three amino acids of this peptide segment was used to affinity purify the fusion proteins in...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1989-06-01 00:00:00
abstract::Automated DNA sequencing requires the intensive use of computers to handle the large amount of data taken. When a computer failure occurs and the data are no longer accessible, all the expense and effort that went into the sequencing experiment is lost. By using the data storage architecture of Macintosh computers to ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/97226bc05
更新日期:1997-06-01 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
pub_type: 杂志文章
doi:10.2144/000113218
更新日期:2009-09-01 00:00:00
abstract::Gene expression profiles may offer more or additional information than classic morphologic- and histologic-based tumor classification systems. Because the number of tissue samples examined is usually much smaller than the number of genes examined, efficient data reduction and analysis methods are critical. In this rep...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/00296bc02
更新日期:2000-12-01 00:00:00
abstract::beta-galactosidase (beta-gal), the product of the E. coli LacZ gene, has been used extensively as a reporter in numerous systems. Until recently, the most commonly used method of detecting beta-gal reporter enzymatic activity was a colormetric assay based on the cleavage of the beta-gal substrate 5-bromo-4-chloro-3-in...
journal_title:BioTechniques
pub_type:
doi:10.2144/01304st03
更新日期:2001-04-01 00:00:00