Fluorescent protein engineering by in vivo site-directed mutagenesis.

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 Life

journal_title

IUBMB life

authors

Valledor M,Hu Q,Schiller P,Myers RS

doi

10.1002/iub.1041

subject

Has Abstract

pub_date

2012-08-01 00:00:00

pages

684-9

issue

8

eissn

1521-6543

issn

1521-6551

journal_volume

64

pub_type

杂志文章
  • Mutational analysis of the three conserved valine residues of insulin and a proposal of "isosteric residue".

    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

    authors: Guo ZY,Tang YH,Zhang Z,Feng YM

    更新日期:2001-12-01 00:00:00

  • Effect of ribozyme against transforming growth factorbeta1 on biological character of activated HSCs.

    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

    authors: Song YH,Zhou XM,Xue XN,Liu NZ,Tian DA,Kong XJ,Wu XL,Lin JS,Jin YX

    更新日期:2005-01-01 00:00:00

  • LncRNA UCA1 impacts cell proliferation, invasion, and migration of pancreatic cancer through regulating miR-96/FOXO3.

    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

    authors: Zhou Y,Chen Y,Ding W,Hua Z,Wang L,Zhu Y,Qian H,Dai T

    更新日期:2018-04-01 00:00:00

  • Porcine induced pluripotent stem cells may bridge the gap between mouse and human iPS.

    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

    authors: Esteban MA,Peng M,Deli Z,Cai J,Yang J,Xu J,Lai L,Pei D

    更新日期:2010-04-01 00:00:00

  • Emerging roles of the αC-β4 loop in protein kinase structure, function, evolution, and disease.

    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

    authors: Yeung W,Ruan Z,Kannan N

    更新日期:2020-06-01 00:00:00

  • Adenylyl cyclase expression and regulation during the differentiation of Dictyostelium discoideum.

    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

    authors: Kriebel PW,Parent CA

    更新日期:2004-09-01 00:00:00

  • Macromolecular mimicry of nucleic acid and protein.

    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

    authors: Pedersen GN,Nyborg J,Clark BF

    更新日期:1999-07-01 00:00:00

  • Role of the SNK-SPAR pathway in the development of Alzheimer's disease.

    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

    authors: Gong X,Lu X,Zhan L,Sui H,Qi X,Ji Z,Niu X,Liu L

    更新日期:2010-03-01 00:00:00

  • Rate of 14CO2 production from variously labeled forms of [14C]glucose in human breast invasive ductal carcinoma tissues.

    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

    authors: Yan SH,Wallon J,Mayaudon J

    更新日期:1999-10-01 00:00:00

  • Characterization of COBRA1 in human breast cancer cell lines using a new polyclonal antibody against COBRA1.

    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

    authors: Zhu J,Song S,Jiang Z,Yan J,Lu Q,Huang C,Ye Q

    更新日期:2004-03-01 00:00:00

  • Conformational stability and thermodynamic characterization of homotetrameric Plasmodium falciparum beta-ketoacyl-ACP reductase.

    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

    authors: Karmodiya K,Sajad S,Sinha S,Maity K,Suguna K,Surolia N

    更新日期:2007-07-01 00:00:00

  • Tea bioactive components prevent carcinogenesis via anti-pathogen, anti-inflammation, and cell survival pathways.

    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

    authors: Fan X,Xiao X,Mao X,Chen D,Yu B,Wang J,Yan H

    更新日期:2021-02-01 00:00:00

  • Paeonol inhibits B16F10 melanoma metastasis in vitro and in vivo via disrupting proinflammatory cytokines-mediated NF-κB and STAT3 pathways.

    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

    authors: Zhang L,Tao L,Shi T,Zhang F,Sheng X,Cao Y,Zheng S,Wang A,Qian W,Jiang L,Lu Y

    更新日期:2015-10-01 00:00:00

  • Impairment of homeostasis in lysosomal storage disorders.

    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

    authors: Segatori L

    更新日期:2014-07-01 00:00:00

  • LAMC1 mRNA promotes malignancy of hepatocellular carcinoma cells by competing for MicroRNA-124 binding with CD151.

    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

    authors: Yang ZP,Ma HS,Wang SS,Wang L,Liu T

    更新日期:2017-08-01 00:00:00

  • FGF receptor mutations: dimerization syndromes, cell growth suppression, and animal models.

    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

    authors: Kannan K,Givol D

    更新日期:2000-03-01 00:00:00

  • miR-200a modulate HUVECs viability and migration.

    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

    authors: Li YX,Liu DQ,Zheng C,Zheng SQ,Liu M,Li X,Tang H

    更新日期:2011-07-01 00:00:00

  • Prostate specific antigen reduction following statin therapy: Mechanism of action and review of the literature.

    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

    authors: Mener DJ

    更新日期:2010-08-01 00:00:00

  • Yeast models of human mitochondrial diseases.

    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

    authors: Barrientos A

    更新日期:2003-02-01 00:00:00

  • Ironing out disease: inherited disorders of iron homeostasis.

    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

    authors: Anderson GJ

    更新日期:2001-01-01 00:00:00

  • Aberrant methylation of APC and RARβ2 genes in breast cancer patients.

    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

    authors: Swellam M,Abdelmaksoud MD,Sayed Mahmoud M,Ramadan A,Abdel-Moneem W,Hefny MM

    更新日期:2015-01-01 00:00:00

  • Interplay of Cu,Zn superoxide dismutase and nitric oxide synthase in neurodegenerative processes.

    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

    authors: Rotilio G,Aquilano K,Ciriolo MR

    更新日期:2003-10-01 00:00:00

  • Search for difference in aminoacylation of mitochondrial DNA-encoded wild-type and mutant human tRNALeu (UUR).

    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

    authors: Wang ZC,Wang XM,Jin YX,Jiao BH,Xu F,Miao MY,Zhu KJ

    更新日期:2003-03-01 00:00:00

  • Hypoxia-induced alterations of transcriptome and chromatin accessibility in HL-1 cells.

    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

    authors: Wang J,Wang Y,Duan Z,Hu W

    更新日期:2020-08-01 00:00:00

  • Interaction between pancreatic cancer cells and tumor-associated macrophages promotes the invasion of pancreatic cancer cells and the differentiation and migration of macrophages.

    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

    authors: Meng F,Li C,Li W,Gao Z,Guo K,Song S

    更新日期:2014-12-01 00:00:00

  • Association of the glucocorticoid receptor with STAT3, C/EBPbeta, and the hormone-responsive element within the rat haptoglobin gene promoter during the acute phase response.

    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

    authors: Arambasić J,Poznanović G,Ivanović-Matić S,Bogojević D,Mihailović M,Uskoković A,Grigorov I

    更新日期:2010-03-01 00:00:00

  • SMC proteins at the crossroads of diverse chromosomal processes.

    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

    authors: Jessberger R

    更新日期:2003-12-01 00:00:00

  • Regulation of cardiac autophagy by insulin-like growth factor 1.

    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

    authors: Troncoso R,Díaz-Elizondo J,Espinoza SP,Navarro-Marquez MF,Oyarzún AP,Riquelme JA,Garcia-Carvajal I,Díaz-Araya G,García L,Hill JA,Lavandero S

    更新日期:2013-07-01 00:00:00

  • Ambient pH sensing in filamentous fungi: pitfalls in elucidating regulatory hierarchical signaling networks.

    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

    authors: Rossi A,Cruz AH,Santos RS,Silva PM,Silva EM,Mendes NS,Martinez-Rossi NM

    更新日期:2013-11-01 00:00:00

  • The HCV ARFP/F/core+1 protein: production and functional analysis of an unconventional viral product.

    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

    authors: Vassilaki N,Mavromara P

    更新日期:2009-07-01 00:00:00