Abstract:
:In vivo site-directed mutagenesis by single-stranded deoxyribonucleic acid recombineering is a facile method to change the color of fluorescent proteins (FPs) without cloning. Two different starting alleles of GFP were targeted for mutagenesis: gfpmut3* residing in the Escherichia coli genome and egfp carried by a bacterial/mammalian dual expression lentiviral plasmid vector. Fluorescent protein spectra were shifted by subtle modification of the chromophore region and residues interacting with the chromophore of the FP. Eight different FPs (Violeta, Azure, Aqua, Mar, Celeste, Amarillo, Mostaza, and Bronze) were isolated and shown to be useful in multicolor imaging and flow cytometry of bacteria and transgenic human stem cells. To make in vivo site-directed mutagenesis more efficient, the recombineering method was optimized using the fluorescence change as a sensitive quantitative assay for recombination. A set of rules to simplify mutant isolation by recombineering is provided.
journal_name
IUBMB Lifejournal_title
IUBMB lifeauthors
Valledor M,Hu Q,Schiller P,Myers RSdoi
10.1002/iub.1041subject
Has Abstractpub_date
2012-08-01 00:00:00pages
684-9issue
8eissn
1521-6543issn
1521-6551journal_volume
64pub_type
杂志文章相关文献
IUBMB LIFE文献大全abstract::To further understand the role of the three conserved Val residues in insulin, B12Val, B18Val, and A3Val, five insulin mutants-[A3Ser]insulin, [B12Thr]insulin, (desB30)[B12Ser]insulin, [B18Thr] insulin, and [B18Leu]insulin--were obtained by means of site-directed mutagenesis and their receptor-binding activities as we...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1080/152165401317291165
更新日期:2001-12-01 00:00:00
abstract::Transforming growth factorbeta1 (TGFbeta1) is considered to be the principal contributor to liver fibrosis. So in this study the ribozymes against TGFbeta1 were designed. The in vitro cleavage activities of the ribozymes were assayed through incubation of (32)p-labeled target RNAs and (32)p-labeled ribozymes in differ...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1080/15216540400024470
更新日期:2005-01-01 00:00:00
abstract::This study was expected to reveal the regulatory effects of lncRNA UCA1 on pancreatic cancer cell progression through targeting miR-96/FOXO3. Microarray analysis was carried out on 36 cases of pancreatic cancer tissues and 16 cases of adjacent tissues among them. Expression levels of lncRNA UCA1, miR-96, and FOXO3 in ...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1002/iub.1699
更新日期:2018-04-01 00:00:00
abstract::Recently, three independent laboratories reported the generation of induced pluripotent stem cells (iPSCs) from pig (Sus scrofa). This finding sums to the growing list of species (mouse, human, monkey, and rat, in this order) for which successful reprogramming using exogenous factors has been achieved, and multiple ot...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1002/iub.307
更新日期:2010-04-01 00:00:00
abstract::The faithful propagation of cellular signals in most organisms relies on the coordinated functions of a large family of protein kinases that share a conserved catalytic domain. The catalytic domain is a dynamic scaffold that undergoes large conformational changes upon activation. Most of these conformational changes, ...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1002/iub.2253
更新日期:2020-06-01 00:00:00
abstract::Cyclic AMP metabolism is essential for the survival of the social amoebae Dictyostelium discoideum. Three distinct adenylyl cyclases are expressed and required for the normal development of this simple eukaryote. The adenylyl cyclase expressed during aggregation, ACA, is related to the mammalian and Drosophila G prote...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1080/15216540400013887
更新日期:2004-09-01 00:00:00
abstract::Although proteins and nucleic acids consist of different chemical components, proteins can mimic structures and possibly also functions of nucleic acids. Recently, structural mimicry was observed between two elongation factors in bacterial protein biosynthesis leading to the introduction of the concept of macromolecul...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1080/713803471
更新日期:1999-07-01 00:00:00
abstract::Alzheimer's disease (AD) is characterized by the presence of senile plaques and neurofibrillary tangles in the brain. The beta-amyloid peptide (Abeta) is the primary constituent of the senile plaques, and has been proposed to be a key contributor to the neurodegeneration observed in AD. The molecular mechanisms underl...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1002/iub.308
更新日期:2010-03-01 00:00:00
abstract::To establish possible cancerous aggressiveness between the metabolism of variously labeled [14C]glucose in the human breast invasive ductal carcinoma (IDC) tissues, we measured the rates of 14CO2 production from those tissues by using radiorespirometry, expressing the results as initial velocity (V) in nanomoles of 14...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1080/713803528
更新日期:1999-10-01 00:00:00
abstract::Mutations in the breast cancer susceptibility gene BRCA1 predispose individuals to breast and ovarian cancers. Cofactor of BRCA1 (COBRA1), a novel protein, was isolated as a BRCA1-interacting protein. However, the role of COBRA1 in breast cancer is poorly understood. In this study, we demonstrate that COBRA1 mRNA was ...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1080/15216540410001699826
更新日期:2004-03-01 00:00:00
abstract::The conformational stability of the homotetrameric Plasmodium falciparum beta-ketoacyl-ACP reductase (FabG) was determined by guanidinium chloride-induced isothermal and thermal denaturation. The reversible unfolding transitions were monitored by intrinsic fluorescence, circular dichroism (CD) spectroscopy and by meas...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1080/15216540701472113
更新日期:2007-07-01 00:00:00
abstract::Cancer seriously impairs human health and survival. Many perturbations, such as increased oxidative stress, pathogen infection, and inflammation, promote the accumulation of DNA mutations, and ultimately lead to carcinogenesis. Tea is one of the most highly consumed beverages worldwide and has been linked to improveme...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1002/iub.2445
更新日期:2021-02-01 00:00:00
abstract::Cancer related inflammation (CRI) is now recognized as the seventh hallmark in the pathogenesis of many types of malignancies. Paeonol, a natural phenolic component isolated from the root bark of Paeonia moutan, has significant anti-inflammatory activity. Recently, accumulating body of research has revealed potent ant...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1002/iub.1435
更新日期:2015-10-01 00:00:00
abstract::Lysosomal storage disorders (LSDs) are inherited metabolic diseases caused by deficiencies in lysosomal proteins, which result in accumulation of undegraded metabolites and disruption of lysosomal proteostasis. Despite significant progress in the molecular genetics and biochemistry underlying the cellular pathogenesis...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1002/iub.1288
更新日期:2014-07-01 00:00:00
abstract::Specific RNAs can function as sinks for endogenous miRNAs, known as competing endogenous RNAs (ceRNAs). Here, we confirm a miR-124 mediated ceRNA crosstalk between LAMC1 and CD151 in hepatocellular carcinoma (HCC). miR-124 negatively regulates LAMC1 expression through two miRNA binding sites within its 3' untranslated...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1002/iub.1642
更新日期:2017-08-01 00:00:00
abstract::This review describes recent progress in the field of fibroblast growth factor receptors (FGFRs) with an emphasis on the role of FGFR mutants in skeletal malformations. This family of four receptors contains the most frequent germline mutations in humans. More than 75 mutations have been recorded, which account for mo...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1080/713803609
更新日期:2000-03-01 00:00:00
abstract::The posttranscriptional regulation of miRNAs is important for organism development. To investigate the role of miRNAs in angiogenesis, we performed a loss-of-function screening assay in human umbilical vein endothelial cells (HUVECs) and found that knockdown of 7 miRNAs (miR-95a, miR-126, miR-129, miR-137, miR-139, mi...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1002/iub.486
更新日期:2011-07-01 00:00:00
abstract::Prostate specific antigen (PSA) is a serine protease that is exclusively produced in the prostate, and its detection is the only laboratory test available for screening men for prostate cancer (PC). The interpretation of the assay is difficult since it is specific for prostate tissue and cellular growth, but not for P...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1002/iub.355
更新日期:2010-08-01 00:00:00
abstract::The yeast Saccharomyces cerevisiae is an excellent model for gaining insights into the molecular basis of human mitochondrial disorders, particularly those resulting from impaired mitochondrial metabolism. Yeast is a very well characterized system and most of our current knowledge about mitochondrial biogenesis in hum...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1002/tbmb.718540876
更新日期:2003-02-01 00:00:00
abstract::Iron presents us with a paradox. Without it, cells simply cannot survive because iron is an essential cofactor for many enzymes in critical biochemical pathways. However, when iron is present in excess, it can be highly cytotoxic due to its propensity to catalyze the formation of reactive oxygen radicals. To cater for...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1080/15216540120450
更新日期:2001-01-01 00:00:00
abstract::Changes in the status of DNA methylation are one of the most common molecular alterations in human neoplasia. We aimed to identify epigenetic molecular markers in serum for early detection of breast cancer. Authors analyzed retrospectively the methylation status of RARβ2 and APC genes in serum samples from 121 breast ...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1002/iub.1346
更新日期:2015-01-01 00:00:00
abstract::Reactive oxygen and nitrogen species (ROS and RNS) have been extensively recognized as important signaling molecules implicated in physiological processes such as gene expression, cell differentiation and immune activation. Nevertheless, continuous production of these species may produce oxidative and/or nitrosative s...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1080/15216540310001628717
更新日期:2003-10-01 00:00:00
abstract::The pathogenetic mechanism of the most extensively investigated A3243G mutated tRNALeu(UUR) gene, which causes the MELAS encephalomyopathy, maternally inherited diabetes, or chronic progressive external ophlthalmoplegia, is still unresolved, despite the numerous investigations on the topic. Previous evidences presente...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1080/1521654031000110190
更新日期:2003-03-01 00:00:00
abstract::Cardiac hypoxia plays a significant role in various types of heart disease, and improper treatment of hypoxia often leads to myocardial cell damage or even death. Transcriptome profiling and open chromatin mapping have been used as powerful tools to understand the development of heart disease, but the interplay betwee...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1002/iub.2297
更新日期:2020-08-01 00:00:00
abstract::In this study, the impact of pancreatic cancer cell interaction with macrophages on the differentiation and function of macrophages and the behaviors of pancreatic cancer cells in vitro is evaluated. The expression of immunocompetent cell-associated markers in 22 pancreatic cancer specimens was characterized by immuno...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1002/iub.1336
更新日期:2014-12-01 00:00:00
abstract::Upregulation of haptoglobin (Hp) expression in the rat during the acute phase (AP) response is the result of synergistic effects of IL-6-, IL-1beta-, and corticosterone-activated signaling pathways. IL-6 signaling terminates in cis-trans interactions of the Hp gene hormone-responsive element (HRE) with transcription f...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1002/iub.313
更新日期:2010-03-01 00:00:00
abstract::How should a protein be designed to serve in processes as diverse as chromosome condensation, sister chromatid cohesion, DNA recombination, gene dosage regulation, and perhaps even gene silencing or transcriptional regulation--which occur in both mitosis and meiosis? Such a protein or protein complex needs to bear DNA...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1080/15216540310001639661
更新日期:2003-12-01 00:00:00
abstract::Insulin-like growth factor-1 (IGF-1) signaling is a key pathway in the control of cell growth and survival. Three critical nodes in the IGF-1 signaling pathway have been described in cardiomyocytes: protein kinase Akt/mammalian target of rapamycin (mTOR), Ras/Raf/extracellular signal-regulated kinase (ERK), and phosph...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1002/iub.1172
更新日期:2013-07-01 00:00:00
abstract::In this article, the experiments used to construct the ambient pH-signaling network involved in the secretion of enzymes by filamentous fungi have been reviewed, focusing on the phosphate-repressible phosphatases in Aspergillus nidulans. Classic and molecular genetics have been used to demonstrate that proteolysis of ...
journal_title:IUBMB life
pub_type: 杂志文章
doi:10.1002/iub.1217
更新日期:2013-11-01 00:00:00
abstract::Hepatitis C virus (HCV) is an enveloped positive-strand RNA virus of the Flaviviridae family. It has a genome of about 9,600 nucleotides encoding a large polyprotein (about 3,000 amino acids) that is processed by cellular and viral proteases into at least 10 structural and nonstructural viral proteins. A novel HCV pro...
journal_title:IUBMB life
pub_type: 杂志文章,评审
doi:10.1002/iub.201
更新日期:2009-07-01 00:00:00