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1.
Article in English | WPRIM | ID: wpr-247126

ABSTRACT

<p><b>OBJECTIVE</b>The complex of the cyclic AMP receptor protein (CRP) and cAMP is an important transcriptional regulator of numerous genes in prokaryotes. The transport of mannitol through the phosphotransferase systems (PTS) is regulated by the CRP-cAMP complex. The aim of the study is to investigate how the CRP-cAMP complex acting on the mannitol PTS operon mtl of the Vibrio cholerae El Tor biotype.</p><p><b>METHODS</b>The crp mutant strain was generated by homologous recombination to assess the need of CRP to activate the mannitol PTS operon of V. cholerae El Tor. Electrophoretic mobility shift assays (EMSA) and the reporter plasmid pBBRlux were used to confirm the role that the CRP-cAMP complex playing on the mannitol PTS operon mtl.</p><p><b>RESULTS</b>In this study, we confirmed that CRP is strictly needed for the activation of the mtl operon. We further experimentally identified five CRP binding sites within the promoter region upstream of the mannitol PTS operon mtl of the Vibrio cholerae El Tor biotype and found that these sites display different affinities for CRP and provide different contributions to the activation of the operon.</p><p><b>CONCLUSION</b>The five binding sites collectively confer the strong activation of mannitol transfer by CRP in V. cholerae, indicating an elaborate and subtle CRP activation mechanism.</p>


Subject(s)
Bacterial Proteins , Genetics , Base Sequence , Cyclic AMP , Metabolism , Cyclic AMP Receptor Protein , Gene Expression Regulation, Bacterial , Mannitol , Molecular Sequence Data , Operon , Phosphotransferases , Vibrio cholerae
2.
Chinese Journal of Epidemiology ; (12): 1105-1110, 2013.
Article in Chinese | WPRIM | ID: wpr-321712

ABSTRACT

<p><b>OBJECTIVE</b>To evaluated the fundamental role of stage control technology (SCT) on the detectability for Salmonella networking laboratories.</p><p><b>METHODS</b>Appropriate Salmonella detection methods after key point control being evaluated, were establishment and optimized. Our training and evaluation networking laboratories participated in the World Health Organization-Global Salmonella Surveillance Project (WHO-GSS) and China-U.S. Collaborative Program on Emerging and Re-emerging infectious diseases Project (GFN) in Shanghai. Staff members from the Yunnan Yuxi city Center for Disease Control and Prevention were trained on Salmonella isolation from diarrhea specimens. Data on annual Salmonella positive rates was collected from the provincial-level monitoring sites to be part of the GSS and GFN projects from 2006 to 2012.</p><p><b>RESULTS</b>The methodology was designed based on the conventional detection procedure of Salmonella which involved the processes as enrichment, isolation, species identification and sero-typing. These methods were simultaneously used to satisfy the sensitivity requirements on non-typhoid Salmonella detection for networking laboratories. Public Health Laboratories in Shanghai had developed from 5 in 2006 to 9 in 2011, and Clinical laboratories from 8 to 22. Number of clinical isolates, including typhoid and non-typhoid Salmonella increased from 196 in 2006 to 1442 in 2011. The positive rate of Salmonella isolated from the clinical diarrhea cases was 2.4% in Yuxi county, in 2012. At present, three other provincial monitoring sites were using the SBG technique as selectivity enrichment broth for Salmonella isolation, with Shanghai having the most stable positive baseline.</p><p><b>CONCLUSION</b>The method of SCT was proved the premise of the network laboratory construction. Based on this, the improvement of precise phenotypic identification and molecular typing capabilities could reach the level equivalent to the national networking laboratory.</p>


Subject(s)
Bacteriological Techniques , Computer Communication Networks , Laboratories , Salmonella , Technology Assessment, Biomedical
3.
Chinese Journal of Epidemiology ; (12): 871-874, 2006.
Article in Chinese | WPRIM | ID: wpr-261720

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the molecular types of Salmonella paratyphi A strains isolated in the recent years, and to construct the standard S. paratyphi A databank in the laboratory surveillance network PulseNet China.</p><p><b>METHODS</b>S. paratyphi A isolates from 4 provinces were analyzed with the standard pulsed-field gel electrophoresis (PFGE) protocol used in PulseNet and their patterns compared. The databank was constructed with BioNumerics.</p><p><b>RESULTS</b>Eleven PFGE patterns were obtained, in which 3 predominant patterns were identifies with a similarity coefficient of 96.3%. The strains of these patterns, accounted for 86.5% of the analyzed strains, appeared in different provinces and years.</p><p><b>CONCLUSION</b>The databank of S. paratyphi A was constructed and could be used in laboratory surveillance of S. paratyphi A in PulseNet China. From the analyses on molecular typing of the isolates, data suggested that the predominant strains might cause the epidemics in different regions.</p>


Subject(s)
China , Electrophoresis, Gel, Pulsed-Field , Salmonella paratyphi A , Classification , Genetics
4.
Article in Chinese | WPRIM | ID: wpr-232131

ABSTRACT

<p><b>OBJECTIVE</b>To study the prevalence of severe acute respiratory syndrome coronavirus (SARS-CoV) like virus in animals at a live animal market of Guanzhou in 2004 before and after culling of wild animal action taken by the local authority, in order to predict the re-emerging of SARS from animal originals in this region.</p><p><b>METHODS</b>Animals at live animal market were sampled for rectal and throat swabs in triplicate. A single step realtime reverse transcription-polymerase chain reaction (RT-PCR) diagnostic kit was performed for screening SARS-CoV like virus, the manual nested RT- PCR and DNA sequencing were performed for confirmation. Only specimens which tested positive for both of the N and P genes by nested RT-PCR were scored as positive.</p><p><b>RESULTS</b>In 31 animals sampled in January 5 2004 before culling of wild animals at Guangdong Province, including 20 cats (Felis catus), 5 red fox (Vulpes vulpes) and 6 Lesser rice field rats (Rattus losea), 8 (25.8%) animals were tested positive for SARS-CoV like virus by RT-PCR methods, of which 4 cats, 3 red fox and one Lesser rice field rats were included. However, two weeks after culling of animals and disinfection of the market were implemented, in 119 animals sampled in January 20 2004, including 6 rabbits (Oryctolagus cuniculus), 13 cats, 46 red jungle fowl (Gallus gallus), 13 spotbill duck (Anas platyrhynchos), 10 greylag goose (Anser anser), 31 Chinese francolin (Franclinus pintadeanus), only rectal swab from one greylag goose was tested positive for SARS-CoV like virus. Furthermore, in 102 animals that including 14 greylag gooses, 3 cats, 5 rabbits, 9 spotbill duck (Anaspoecilorhyncha), 2 Chinese francolin (Franclinus pintadeanus), 8 common pheasant (Phasianus colchicus), 6 pigeons, 9 Chinese muntjac (Muntiacus reevesi), 19 wild boar (Sus scrofa), 16 Lesser rice field rats, 5 dogs, 1 mink (Mustela vison), 3 goats, 2 green peafowl (Pavo muticus) sampled in April, May, June, July, August and November, only rectal swab from one pig was tested positive. However, of 12 and 10 palm civets sampled in November and December including five of which had been at the live animals market for 2 days, none of them was tested positive.</p><p><b>CONCLUSION</b>This findings revealed that animals being sampled in April, May, June, July, August and November of 2004, only one rectal swab from a pig was tested positive as SARS-CoV like virus, much lower than the results from the previous year, suggesting that the possibility of re-emerging of human infection from animal origins is low for the winter of 2004-2005.</p>


Subject(s)
Animals , Animals, Wild , Virology , China , DNA, Viral , Felidae , Virology , Reverse Transcriptase Polymerase Chain Reaction , Severe acute respiratory syndrome-related coronavirus
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