Consulta: subjectFacets:"Molecular cloning"
Registros recuperados: 14
Data/hora: 06/06/2024 10:31:02

Expression of a Haemonchus contortus cysteine protease in the baculovirus system
Provedor de dados: 69
Autores: Miranda-Miranda,Estefan; Murillo-Sánchez,María Hortensia; Cossío-Bayúgar,Raquel.
A Haemonchus contortus recombinant Cysteine Protease (CP) was expressed in the baculovirus system. The CP gene was isolated by PCR from H. contortus cDNA, the PCR amplicon was cloned downstream to the polihedrin promoter within a bacterial expression vector, Sf9 insect cells were used for simultaneous co-transfection with the CP-vector and baculovirus naked DNA, which originated recombinant viruses by homologous recombination capable to express recombinant CP in an insect cell culture. A recombinant protease was identified as a fusion protein with a Ni lithium affinity 6XHis group. Recombinant CP was purified by affinity chromatography to obtain active recombinant protease identified by H. contortus experimentally infested ovine sera on a western blot as a...
Tipo: Journal article
Palavras-chave: Haemonchosis; Molecular cloning; Recombinant protease; Synthetic substrates; Western blot.
Ano: 2008
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000200007

Development and significance of RAPD-SCAR markers for the identification of Litchi chinensis Sonn: by improved RAPD amplification and molecular cloning
Provedor de dados: 69
Autores: Cheng,Jingliang; Long,Yan; Khan,Asaduzzaman; Wei,Chunli; Fu,Shelly; Fu,Junjiang.
Background Analysis of genetic diversity is important for the authentication of a species. Litchi (Litchi chinensis Sonn.) is a subtropical evergreen tree. Recently, L. chinensis has been characterized by an improved random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) analysis. The goal of this study was to develop sequence-characterized amplified region (SCAR) markers from the improved RAPD fragments for the genetic analysis of L. chinensis. Results The improved RAPD fragments from L. chinensis were cloned, sequenced and converted into stable SCAR markers. Sequencing of three cloned RAPD fragments revealed that the clone L7-16 consisted of 222 nucleotides (GenBank accession number KM235222), clone L9-6 consisted of 648...
Tipo: Journal article
Palavras-chave: Genetic authentication; Molecular cloning; Random amplified polymorphic DNA; Sequence-characterized amplified region marker.
Ano: 2015
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582015000100007

Molecular cloning and in silico analysis of the duck (Anas platyrhynchos) MEF2A gene cDNA and its expression profile in muscle tissues during fetal development
Provedor de dados: 74
Autores: Liu,Hehe; Wang,Jiwen; Si,Jianmin; Jia,Jing; Li,Liang; Han,Chunchun; Huang,Kailiang; He,Hua; Xu,Feng.
The role of myogenic enhancer transcription factor 2a (MEF2A) in avian muscle during fetal development is unknown. In this work, we cloned the duck MEF2A cDNA sequence (GenBank accession no. HM460752) and examined its developmental expression profiles in cardiac muscle, non-vascular smooth muscle and skeletal muscle. Duck MEF2A cDNA comprised 1479 bp encoding 492 amino acid residues. In silico analysis showed that MEF2A contained MADS (MCM1, AGAMOUS, DEFICIENS and SRF -serum response factor), MEF2 and mitogen-activated protein kinase (MAPK) transcription domains with high homology to related proteins in other species. Modified sites in these domains were conserved among species and several variants were found. Quantitative PCR showed that MEF2A was...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Duck MEF2A; Expression profile; In silico analysis; Molecular cloning; Muscle tissues.
Ano: 2012
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000100026

Molecular cloning and expression profiling of a chalcone synthase gene from hairy root cultures of Scutellaria viscidula Bunge
Provedor de dados: 74
Autores: Lei,Wei; Tang,Shao-Hu; Luo,Ke-Ming; Sun,Min.
A cDNA encoding chalcone synthase (CHS), the key enzyme in flavonoid biosynthesis, was isolated from hairy root cultures of Scutellaria viscidula Bunge by rapid amplification of cDNA ends (RACE). The full-length cDNA of S. viscidula CHS, designated as Svchs (GenBank accession no. EU386767), was 1649 bp with a 1170 bp open reading frame (ORF) that corresponded to a deduced protein of 390 amino acid residues, a calculated molecular mass of 42.56 kDa and a theoretical isoelectric point (pI) of 5.79. Multiple sequence alignments showed that SvCHS shared high homology with CHS from other plants. Functional analysis in silico indicated that SvCHS was a hydrophilic protein most likely associated with intermediate metabolism. The active sites of the malonyl-CoA...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Chalcone synthase gene; Methyl jasmonate; Molecular cloning; Scutellaria viscidula Bunge.
Ano: 2010
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572010000200015

Molecular characterization of penaeidins from two Atlantic Brazilian shrimp species, Farfantepenaeus paulensis and Litopenaeus schmitti
Provedor de dados: 5
Autores: Barracco, Margherita; De Lorgeril, Julien; Gueguen, Yannick; Bachere, Evelyne.
We report here the molecular cloning of new members of the penaeidin family from two Atlantic penaeids from Brazil, Litopenaeus schmitti and Farfantepenaeus paulensis. The presence of penaeidins in the granular hemocytes of both shrimps was first evidenced by immunofluorescence, using polyclonal antibodies raised against L. vannamei penaeidin Litvan PEN3-1. cDNAs from the hemocytes of both Brazilian species were obtained by reverse transcription and the sequences encoding penaeidins were amplified by PCR, using primers based on penaeidin consensus sequences. Five penaeidin clones were obtained. According to the international penaeidin classification (PenBase, http://www.penbase.immunaqua.com), the deduced amino acid sequences of two clones from L. schmitti...
Tipo: Text
Palavras-chave: Immuno detection; Molecular cloning; Litopenaeus schmitti; Farfantepenaeus paulensis; Brazilian penaeid shrimps; Hemocytes; Antimicrobial peptides; Penaeidins.
Ano: 2005
URL: http://archimer.ifremer.fr/doc/2005/publication-3582.pdf

Heterologous expression of an insecticidal gene from the armed spider (Phoneutria nigriventer)
Provedor de dados: 80
Autores: Figueiredo,J. E. F.; Kalapothakis,E.; Gomez,M. V.; Bressan,W.
Insect-pests are global problems that cause severe damage to crop plants, and their control is commonly based on chemical insecticides. However, negative effects of pesticides on the environment and human health emphasize the necessity to develop alternative methods for insect-pest control. In the present study, a gene coding for the insecticidal peptide TX4(6-1) of the Brazilian armed spider (Phoneutria nigriventer) was cloned in fusion with maltose binding protein (MBP) and expressed in Escherichia coli. The affinity purified protein MBP-GlyTX4 was cleaved with the Xa factor and used for a bioassay against Spodoptera frugiperda and rabbit immunization. Five micrograms GlyTX4 protein injected into the hemocoel of larvae and abdominal cavity of adults...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Bioinsecticide; Molecular cloning; Heterologous expression; Escherichia coli; Spodoptera frugiperda.
Ano: 2008
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1678-91992008000200006

An effective method for cloning of partial MADS-box genes related to flower development in groundnut
Provedor de dados: 30
Autores: Yuan, M.
Palavras-chave: Genes; PCR; DNA; Cloning; Transcription factors; Genomes; Transcription; Genetic structures; Nucleotides; Molecular cloning.
Ano: 2005
URL: http://agropedia.iitk.ac.in/openaccess/?q=node/3213

Paternity test in "Mangalarga-Marchador" equines by DNA-fingerprinting
Provedor de dados: 86
Autores: ANUNCIAÇÃO,CARLOS EDUARDO; ASTOLFI-FILHO,SPARTACO.
GC-rich molecular minisatellite probes isolated from the human genome have presented a poor ability for individualization in horses. In this study new DNA sequences were isolated which could be used in paternity tests in horses. Genomic DNA from "Mangalarga-Marchador" horses was treated with restriction enzymes that preferentially digest non-repetitive sequences, so preserving the structure where mini and microsatellites are located. Four clones (S01, S05, S07 and S09) selected from a genomic library screened with a (TG)n oligonucleotide showed similar hybridization profiles generating bands of DNA-fingerprinting type. Using these probes the individualization power obtained was 10-8, which is 10(5)fold higher than that obtained with M13, another GC-rich...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Breeding methods; Molecular cloning; Progeny testing; Horses; Identification; Genetic polymorphism.
Ano: 2000
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-204X2000001000012

Laboratory Protocols 2008: Maize Nutrition Quality and Plant Tissue Analysis Laboratory
Provedor de dados: 31
Autores: Gallicia,   L.; Nurit,   E.; Rosales,   A.; Palacios-Rojas,   N.
Tipo: Report
Palavras-chave: Zea mays; Maize; Wheats; Triticum; Transgenic plants; Genetic engineering; Gene transfer; Genetic transformation; DNA hybridization; Phenotypes; Selection; Tissue culture; In vitro culture; Biolistics; Research projects; Methods; Molecular cloning; PCR; Crop Production/Industries; F02; F30.
Ano: 2009
URL: http://purl.umn.edu/56177

Molecular Cloning and Mrna Expression Analysis of Sichuan White Goose (Anser Cygnoides) Chrebp Gene
Provedor de dados: 89
Autores: Xu,HY; Tang,H; Pan,ZX; Li,L; Han,CC; Liu,HH; He,H; Kang,B; Hu,JW; Xia,L; Wang,Y; Wang,JW.
ABSTRACT The carbohydrate response element-binding protein (ChREBP) is an important nuclear factor that regulates glycolysis and de novo lipogenesis. However, the role of ChREBP in fatty liver development in geese remains unclear. In order to understand the function of ChREBP in lipid metabolism of geese, we first cloned the complete cDNA of the ChREBP of the Sichuan White goose (Anser cygnoides) using RT-PCR, 5’ RACE and 3’ RACE, and analyzed goose ChREBP expression in nine different tissues using real-time PCR technology. The results showed that the goose ChREBP CDS consists of 945bp nucleotides that encode 314 amino acids, and the sequence has high similarities with the swan goose (Anser cygnoides domesticus) and duck (Anas platyrhynchos) sequences,...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: ChREBP; Expression profile; Molecular cloning; RACE; Sichuan White Goose.
Ano: 2017
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-635X2017000400615

A two-plasmid strategy for engineering a dengue virus type 3 infectious clone from primary Brazilian isolate
Provedor de dados: 42
Autores: SANTOS,JEFFERSON J.S.; CORDEIRO,MARLI T.; BERTANI,GIOVANI R.; MARQUES,ERNESTO T.A.; GIL,LAURA H.V.G.
Dengue infections represent one of the most prevalent arthropod-borne diseases worldwide, causing a wide spectrum of clinical outcomes. Engineered infectious clone is an important tool to study Dengue virus (DENV) biology. Functional full-length cDNA clones have been constructed for many positive-strand RNA viruses and have provided valuable tools for studying the molecular mechanisms involved in viral genome replication, virion assembly, virus pathogenesis and vaccine development. We report herein the successful development of an infectious clone from a primary Brazilian isolate of dengue virus 3 (DENV3) of the genotype III. Using a two-plasmid strategy, DENV3 genome was divided in two parts and cloned separately into a yeast-bacteria shuttle vector. All...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Reverse genetics; Dengue virus; Molecular cloning; Infectious clone.
Ano: 2014
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652014000401749

Cloning and Characterization of the Gene Encoding 3-hydroxy-3- Methylglutaryl-coenzyme A (HMG-CoA) Reductase from Fritillaria Cirrhosa D. Don
Provedor de dados: 52
Autores: Zhao,Qi; Li,Rui; Chen,Xiao; Yang,Qian; Li,Jian.
ABSTRACT The enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR; EC1.1.1.34) catalyzes the first committed step of isoprenoids biosynthesis in Mevalonate (MVA) pathway. Here we report for the first time the cloning and characterization of a full-length cDNA encoding HMGR from Fritillaria cirrhosa (FcHMGR), a bulbous medicinal plant. The full-length cDNA of FcHMGR was 2072 base pair (bp), containing a 1680-bp open reading frame. Bioinformatical analyses revealed that FcHMGR had HMG CoA-binding domains and two NADPH binding domains, which are required for HMGR activity. Quantitative real-time PCR (qRT-PCR) analysis revealed that FcHMGR expressed high in mature bulbs. A truncated version of FcHMGR protein lacking the N-terminal 249-bp GC rich area was...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: 3-Hydroxy-3-methylglutaryl-CoA reductases; Fritillaria cirrhosa; Molecular cloning; Expression pattern; Prokaryotic expression.
Ano: 2018
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132018000100438

Analysis of microbial diversity in Shenqu with different fermentation times by PCR-DGGE
Provedor de dados: 58
Autores: Liu,Tengfei; Jia,Tianzhu; Chen,Jiangning; Liu,Xiaoyu; Zhao,Minjie; Liu,Pengpeng.
Abstract Shenqu is a fermented product that is widely used in traditional Chinese medicine (TCM) to treat indigestion; however, the microbial strains in the fermentation process are still unknown. The aim of this study was to investigate microbial diversity in Shenqu using different fermentation time periods. DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) profiles indicated that a strain of Pediococcus acidilactici (band 9) is the predominant bacteria during fermentation and that the predominant fungi were uncultured Rhizopus, Aspergillus oryzae, and Rhizopus oryzae. In addition, pathogenic bacteria, such as Enterobacter cloacae, Klebsiella oxytoca, Erwinia billingiae, and Pantoea vagan were detected in Shenqu. DGGE analysis...
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Microbial diversity; PCR-DGGE; Shenqu; Molecular cloning.
Ano: 2017
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822017000200246

New vectors derives from pUC 18 for clonig and thermal-induced expression in Escherichia coli
Provedor de dados: 58
Autores: Xavier,Mauro Aparecido Souza; Kipnis,André; Torres,Fernando Araripe Gonçalves; Astofi-Filho,Spartaco.
We report the construction of two vectors for Escherichia coli: pUC72, for molecular cloning, and pPLT7, for thermal-induced expression. The main feature of pUC72 is a novel polylinker region that includes restriction sites for Nde I and Nco I which provide an ATG codon for proper translation initiation of expressed genes. Vector pPLT7 is ideal for thermo-inducible expression in host cells that carry the cI857 repressor gene. The use of pPLT7 was validated by the successful expression of the genes encoding carp and porcine growth hormones. These vectors provide novel cloning possibilities in addition to simple, non-expensive, high level expression of recombinant proteins in E. coli.
Tipo: Info:eu-repo/semantics/article
Palavras-chave: Heterologous expression; Escherichia coli; Molecular cloning; Induced expression.
Ano: 2009
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822009000400007