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| MIQUEL,ALVARO; MÜLLER,ILSE; FERRER,PABLO; VALENZUELA,PABLO D. T.; BURZIO,LUIS O. |
| We have used the expression library immunization technology to study the protection of Coho salmon Oncorhynchus kisutch to the infection with Piscirickettsia salmonis. Purified DNA from this bacterium was sonicated and the fragments were cloned in the expression vector pCMV-Bios. Two libraries were obtained containing 22,000 and 28,000 colonies and corresponding to approximately 8 and 10 times the genome of the pathogen, respectively. On average, the size of the inserts ranged between 300 and 1,000 bp. The plasmid DNA isolated from one of these libraries was purified and 20 µg were injected intramuscularly into 60 fish followed by a second dose of 10 µg applied 40 days later. As control, fish were injected with the same amount of DNA of the vector... |
| Tipo: Journal article |
Palavras-chave: Piscirickettsia salmonis; Genetic vaccine; Expression library immunization. |
| Ano: 2003 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602003000300004 |
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| WILHELM,VIVIAN; HUARACÁN,BERNARDO; MARTÍNEZ,RODRIGO; ROSEMBLATT,MARIO; BURZIO,LUIS O; VALENZUELA,PABLO D.T. |
| The genes encoding the heat shock proteins HSP10 and HSP16 of the salmon pathogen Piscirickettsia salmonis have been isolated and sequenced. The HSP10 coding sequence is located in an open reading frame of 291 base pairs encoding 96 aminoacids. The HSP16 coding region was isolated as a 471 base pair fragment encoding a protein of 156 aminoacids. The deduced aminoacid sequences of both proteins show a significant homology to the respective protein from other prokaryotic organisms. Both proteins were expressed in E. coli as fusion proteins with thioredoxin and purified by chromatography on Ni-column. A rabbit serum against P. salmonis total proteins reacts with the recombinant HSP10 and HSP16 proteins. Similar reactivity was determined by ELISA using serum... |
| Tipo: Journal article |
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| Ano: 2003 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602003000300013 |
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