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Heterologous protein secretion in Lactococcus lactis: a novel antigen delivery system BJMBR
Langella,P.; Le Loir,Y..
Lactic acid bacteria (LAB) are Gram-positive bacteria and are generally regarded as safe (GRAS) organisms. Therefore, LAB could be used for heterologous protein secretion and they are good potential candidates as antigen delivery vehicles. To develop such live vaccines, a better control of protein secretion is required. We developed an efficient secretion system in the model LAB, Lactococcus lactis. Staphylococcal nuclease (Nuc) was used as the reporter protein. We first observed that the quantity of secreted Nuc correlated with the copy number of the cloning vector. The nuc gene was cloned on a high-copy number cloning vector and no perturbation of the metabolism of the secreting strain was observed. Replacement of nuc native promoter by a strong...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Protein secretion; Staphylococcal nuclease; Lactococcus lactis; Propeptide; Bovine coronavirus; Epitope.
Ano: 1999 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X1999000200007
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Molecular analysis of the bovine coronavirus S1 gene by direct sequencing of diarrheic fecal specimens BJMBR
Takiuchi,E.; Alfieri,A.F.; Alfieri,A.A..
Bovine coronavirus (BCoV) causes severe diarrhea in newborn calves, is associated with winter dysentery in adult cattle and respiratory infections in calves and feedlot cattle. The BCoV S protein plays a fundamental role in viral attachment and entry into the host cell, and is cleaved into two subunits termed S1 (amino terminal) and S2 (carboxy terminal). The present study describes a strategy for the sequencing of the BCoV S1 gene directly from fecal diarrheic specimens that were previously identified as BCoV positive by RT-PCR assay for N gene detection. A consensus sequence of 2681 nucleotides was obtained through direct sequencing of seven overlapping PCR fragments of the S gene. The samples did not undergo cell culture passage prior to PCR...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bovine coronavirus; S1 gene; Sequencing; Genetic polymorphism.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2008000400004
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Isolation in HRT-18 cells and molecular analysis of a BCoV strain from diarrheic feces of naturally infected calves BABT
Stipp,Danilo Tancler; Barry,Aline Fernandes; Alfieri,Alice Fernandes; Bodnar,Lívia; Alfieri,Amauri Alcindo.
Bovine coronavirus (BCoV) may cause acute diarrhea in newborn calves, leading to significant economic losses for cattle farmers. There are several diagnostic techniques used to detect BCoV in calf fecal samples, but virus isolation still has advantages for antigenic and genomic characterization. This study describes the isolation in HRT-18 cells and molecular characterization of Brazilian BCoV wild-type strains. Three fecal samples from diarrheic 30 day-old calves were inoculated in HRT-18 cell monolayers, which were then evaluated for HA titers and tested using semi-nested PCR followed by RFLP and sequencing. Two samples were successfully isolated and presented HA titers of 16 and 32 units per 25 mL. The results were confirmed using semi-nested PCR and...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cattle; Diarrhea; Bovine coronavirus; Cell culture; Molecular analyses.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000700007
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An outbreak of winter dysentery caused by bovine coronavirus in a high-production dairy cattle herd from a tropical country BABT
Takiuchi,Elisabete; Barry,Aline Fernandes; Alfieri,Alice Fernandes; Filippsen,Patrícia; Alfieri,Amauri Alcindo.
Bovine coronavirus (BCoV) is a known cause of winter dysentery (WD) in adult cattle. The morbidity of the disease is high, that results in a significant decrease in milk production and consequently, economic losses. In the present study, we report on a classical outbreak of WD that affected a high-production Holstein dairy herd raised in a tropical country. The lactating batch included 154 cows, and 138 (90%) presented diarrhea in a short (nine days) period of time. Three (2%) cows died. The other batches of animals did not become ill. The evolution of the disease in the herd, including the clinical signs and epidemiological features, strongly suggested a WD case. Semi-nested PCR and RFLP confirmed that BCoV was the cause of the infection. Samples tested...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cattle; Dairy herd; Winter dysentery; Diarrhea; Bovine coronavirus.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000700008
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Differentiation of Bovine coronavirus (BCoV) genotypes by a restriction enzyme assay BJM
Souza,Sibele Pinheiro de; Asano,Karen Miyuki; Sandri,Thaisa Lucas; Barros,Iracema Nunes de; Richtzenhain,Leonardo José; Brandão,Paulo Eduardo.
This article reports the use of the GsuI restriction enzyme to differentiate genotypes of Bovine Coronavirus (BCoV), based on an 18-nucleotide deletion of S1-coding region found in one of the two genotypes. It was concluded that this assay can be used as a rapid tool for BCoV genotypes differentiation.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bovine coronavirus; Restriction enzyme; Genotypes; Gene S.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822010000300034
Registros recuperados: 5
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