Abstract:
:A method has been developed for the specific amplification and cloning of human hprt cDNA which can be used for mutant sequence analysis. Messenger RNA is isolated from TK6 lymphoblasts and is used to produce a first strand cDNA with reverse transcriptase primed with oligo dT. Second strand synthesis and subsequent amplification of hprt sequences is accomplished using Thermus aquaticus DNA polymerase and hprt-specific primers in the polymerase chain reaction (PCR) procedure. Convenient restriction enzyme sites have been built into the 5' ends of the PCR primers to allow cloning of the hprt fragments in M13mp19. Dideoxy sequencing of hprt with specific primers can be carried out using either the PCR reaction product or fragments cloned in M13mp19 as substrate. This general cloning/sequencing method can be used to analyze hprt mutation in human cells obtained both in vitro and in vivo.
journal_name
Biochem Biophys Res Communjournal_title
Biochemical and biophysical research communicationsauthors
Simpson D,Crosby RM,Skopek TRdoi
10.1016/0006-291x(88)90619-5subject
Has Abstractpub_date
1988-02-29 00:00:00pages
487-92issue
1eissn
0006-291Xissn
1090-2104pii
0006-291X(88)90619-5journal_volume
151pub_type
杂志文章abstract::The growth and metastasis of tumors depend on angiogenesis. Tumor angiogenesis is initiated by the secretion of growth factors from tumor cells; downstream signals are then triggered in pre-existing blood vessels to sprout a new vascular network. Trichosanthin (TCS) is a type I ribosome-inactivating protein that has a...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2012.11.080
更新日期:2013-01-11 00:00:00
abstract::Bacillus thuringiensis var. israelensis produces 130 kDa delta-endotoxin which is highly toxic to mosquito-larvae. The mosquito-larvicidal activity was delineated by sequential deletions from ends of the 1136 amino acids delta-endotoxin. A maximum of 459 amino acids could be removed from the carboxy-terminal of the to...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/s0006-291x(88)81221-x
更新日期:1988-05-31 00:00:00
abstract::We have devised a method to evaluate the capacity of mammalian cell extracts to incise damaged DNA in vitro. The assay uses damaged-plasmid DNA as a substrate for nucleotide excision repair by cell extracts. During this process, enzymatic incision of the damaged DNA is followed by DNA resynthesis. Under our assay cond...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1006/bbrc.1994.1999
更新日期:1994-07-29 00:00:00
abstract::Recombinant baculoviruses expressing full length and 3' truncated forms of c-DNA encoding the Drosophila melanogaster acetylcholinesterase (AChE) were constructed. Biochemical analyses showed that full length recombinant protein was enzymatically active and anchored to the cell membrane via a glycolipidic residue. DTT...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1006/bbrc.1994.2243
更新日期:1994-08-30 00:00:00
abstract::Incubation of purified protein kinase C with H2O2 results in the generation of a persistently activated form of the enzyme which is no longer dependent on Ca2+ or lipid cofactors. This oxidative activation of purified protein kinase C requires added Fe2+ in the incubation medium. Treatment of the soluble fraction of h...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/0006-291x(92)90463-u
更新日期:1992-09-30 00:00:00
abstract::We describe a new approach to DNA hybridization assays using metal-enhanced fluorescence. Thiolated oligonucleotides were bound to silver particles on a glass substrate. Addition of a complementary fluorescein-labeled oligonucleotide resulted in a dramatic time-dependent 12-fold increase in fluorescence intensity duri...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/s0006-291x(03)00935-5
更新日期:2003-06-20 00:00:00
abstract::The human pregnane X receptor (hPXR) plays a key role in the regulation of both drug metabolism and efflux by inducing the expression of CYP3A4 and MDR1 gene. Using reverse transcription-polymerase chain reaction (RT-PCR) analysis, we identified seven novel splicing variants of hPXR in tissue from a single human liver...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/s0006-291x(02)02469-5
更新日期:2002-11-01 00:00:00
abstract::Allylisothiocyanate (ANCS) is shown to inhibit (H+ + K+)-ATPase isolated from the parietal cell of hog gastric mucosa. The ATPase is the proton pump in the secretory membrane of the parietal cell and is an essential component of the acid secretory mechanism. Furthermore, ANCS suppresses acid secretion by bullfrog gast...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/0006-291x(83)91487-0
更新日期:1983-04-29 00:00:00
abstract::Platelet-derived growth factor-D (PDGF-D) is a more recent recognized growth factor involved in the regulation of several cellular processes, including cell proliferation, transformation, invasion, and angiogenesis by binding to and activating its cognate receptor PDGFR-β. After bile duct ligation or in the carbon tet...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2014.12.106
更新日期:2015-02-13 00:00:00
abstract::Prion diseases are characterized by the accumulation of a pathological form of prion protein (PrP(Sc)), which behaves as an infectious agent. Here we developed an in vitro co-culture system to analyze the PrP(Sc) transmission from ScN2a cell, which persistently retains PrP(Sc), to naïve N2a cell. In this cell-to-cell ...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2010.05.143
更新日期:2010-07-02 00:00:00
abstract::Prion diseases are fatal neurodegenerative diseases associated with structural conversion of α-helical prion protein (PrP) into its β-sheet rich isoform (PrPSc). Previous genetic analyses have indicated that several amino acid residues involved in the hydrophobic core of PrP (such as V180, F198, and V210) play a criti...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2018.10.075
更新日期:2018-11-17 00:00:00
abstract::During our human heart cDNA sequencing project, we have obtained a novel cDNA clone which is very similar in DNA and amino acid sequences to a rat/mouse cysteine-rich intestinal protein (1). Sequence analysis has shown that this human cysteine-rich heart protein (hCRHP) is a protein of 77 amino acids and possesses a L...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1006/bbrc.1994.2693
更新日期:1994-11-30 00:00:00
abstract::Focal adhesions (FAs) connect the cellular actin cytoskeleton via integrin with the extracellular matrix. They comprise of many structural and signaling proteins which are highly dynamic, well regulated, and responsible for the sensing of physical properties from the environment. Vinculin is a protein that incorporate...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2010.07.110
更新日期:2010-09-03 00:00:00
abstract::Two isoforms of the src-family tyrosine kinase p59fyn have been shown to arise through alternative splicing of exon 7 of the fyn gene. These isoforms have been designated fynT, expressed in hematopoietic cells, and fynB, expressed in the brain. Here, we describe a novel isoform, fynDelta7, in which exon 7 is absent. F...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/s0006-291x(02)02510-x
更新日期:2002-11-08 00:00:00
abstract::Vaspin, an adipocytokine recently identified in a rat model of type 2 diabetes, has been suggested to have an insulin-sensitizing effect. However, the exact mechanism underlying this action has not been fully elucidated. Furthermore, the specific function of vaspin is largely unknown, especially in vascular cells. We ...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2011.08.083
更新日期:2011-09-23 00:00:00
abstract::Tyrosine protein kinase activity was measured in membranes from DMBA-induced mammary tumors, with Angiotensin II as substrate. The apparent Km for the peptide was 3.3 mM. This enzymatic activity is inhibited by Ca+2; Mn+2 can replace Mg+2 with an increase in the Km for ATP from 47 /microM to 172 microM. The enzymatic ...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/s0006-291x(84)80264-8
更新日期:1984-09-28 00:00:00
abstract::The cDNA encoding Mn peroxidase isozyme H4 from Phanerochaete chrysosporium was recombined into a baculovirus and heterologously expressed in Sf9 cells. The recombinant Mn peroxidase has the same molecular weight as the native enzyme as determined by SDS-PAGE and cross-reacts with a Mn peroxidase-specific antibody. Th...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/0006-291x(91)91903-p
更新日期:1991-09-16 00:00:00
abstract::Kaiso was previously described as a methylated DNA-binding protein and a transcription repressor interacting with the corepressor protein complex NCoR. In the current study, we show that generation-3 Kaiso knockout mice show a phenotype of splenomegaly and large diffused germinal centers (GC). In the spleens of Kaiso ...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2013.11.046
更新日期:2013-12-13 00:00:00
abstract::An antifungal protein, AtUSP protein (At3g53990), was isolated from Arabidopsis thaliana leaves by ion and size chromatography and sequenced by N-terminal sequencing. The AtUSP gene amplified from an Arabidopsis leaf cDNA library was transformed to Escherichia coli to express the AtUSP protein. The recombinant protein...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2017.03.126
更新日期:2017-05-13 00:00:00
abstract::Quercetin is one of the most prominent dietary antioxidants. During its antioxidant activity, quercetin becomes oxidized into its o-quinone/quinone methide QQ. QQ is toxic since it instantaneously reacts with thiols of, e.g., proteins. In cells, QQ will initially form an adduct with glutathione (GSH), giving GSQ. We h...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2005.10.031
更新日期:2005-12-16 00:00:00
abstract::Tuberculosis and HIV continue to be the world-leading killers among infectious diseases, primarily affecting poor people in many developing countries. Despite differences in the immunopathogenesis of human infection with tuberculosis and HIV, experimental evidence from clinical studies and relevant animal models can b...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章,评审
doi:10.1016/j.bbrc.2010.02.141
更新日期:2010-05-21 00:00:00
abstract::We demonstrate that the 204 bp intronic gene fragment of C3(1), which has a specific in vitro affinity for the androgen receptor, is able to confer androgen responsiveness to a heterologous promoter. This characteristic is completely destroyed by a single G----T substitution, affecting a 5'-TGTTCT-3' element that clos...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/0006-291x(89)91534-9
更新日期:1989-10-31 00:00:00
abstract::Rat hearts were perfused for 15min with buffer equilibrated with 0.01% or 0.05% CO. The buffer was equilibrated with 21% O(2) throughout. The ventricular glutathione content decreased by 76% and 84%, 90min post-exposure to 0.01% and 0.05% CO, respectively, compared with 0% CO controls (0.45+/-0.01 micromol/g wet tissu...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/s0006-291x(03)00190-6
更新日期:2003-03-07 00:00:00
abstract::Sensitization to Cupressaceae pollen has become one of the most important causes of pollinosis in Western countries during winter and early spring. However, the characterization of the extracts, the allergens involved and the cross-reactivity with other pollen sources still remain poorly studied; in the case of Cupres...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2010.09.079
更新日期:2010-10-22 00:00:00
abstract::Liposomes prepared from lecithin:cholesterol:p-aminophenyl-alpha-mannoside (7:2:1, v/v/v) were efficiently incorporated into the mouse brain across the blood brain barrier. Furthermore, liposomes injected intraperitoneally were exclusively distributed into lysosome rich fraction and also taken up by glial cells. These...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/s0006-291x(88)81333-0
更新日期:1988-06-30 00:00:00
abstract::Platelets play essential roles in hemostasis and thrombosis by aggregating with each other. However, the molecular mechanism governing platelet aggregation is not yet fully understood. Here, we established an assay system using platelets permeabilized with streptolysin-O to analyze mechanism of the thrombin-induced ag...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1006/bbrc.2001.4237
更新日期:2001-02-02 00:00:00
abstract::Ca2+-phospholipid dependent protein kinase activity and the effect of 12-O-tetradecanoylphorbol-13-acetate (TPA) on H+ secretion was studied in guinea-pig parietal cells. In resting parietal cells PK-C activity was distributed almost equally in both the cytosolic and particulate cell fraction. Exposure of the cells to...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/0006-291x(87)90427-x
更新日期:1987-12-16 00:00:00
abstract::Dictyostelium discoideum possesses more EGF-like (EGFL) domains than any other sequenced eukaryote. Here we show that a synthetic EGFL peptide (DdEGFL1) based upon an amino acid sequence from a cysteine-rich Dictyostelium protein, functions extracellularly to enhance random cell motility and cAMP-mediated chemotaxis i...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2008.12.081
更新日期:2009-02-06 00:00:00
abstract::The ability of various compounds to stimulate cyclic GMP accumulation was studied in neuronal and astrocyte-enriched primary cultures from rat cerebrum. Glutamate was the only agonist eliciting a response in neurons whereas several agonists had an effect in astrocytes but only those due to norepinephrine and glutamate...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/0006-291x(92)91883-r
更新日期:1992-02-14 00:00:00
abstract::S-nitrosothiols (SNOs) are endogenous signaling molecules with a broad spectrum of beneficial airway effects. SNOs are normally present in the airway, but levels tend to be low in cystic fibrosis (CF) patients. We and others have demonstrated that S-nitrosoglutathione (GSNO) increases the expression, maturation, and f...
journal_title:Biochemical and biophysical research communications
pub_type: 杂志文章
doi:10.1016/j.bbrc.2013.12.130
更新日期:2014-01-24 00:00:00