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1.
猪肺炎支原体黏附因子基因R1R2区的克隆及表达   总被引:4,自引:0,他引:4  
根据GenBank登录的猪肺炎支原体232株P97基因和J株黏附因子基因设计了1对引物,以我国猪肺炎支原体Z株(强毒株)基因组DNA为模板,通过PCR方法扩增了该株黏附因子基因的部分序列。经序列分析后,重新设计了1对带有EcoRI和HindⅢ酶切位点的引物,并经引物的定点突变,PCR扩增了Z株黏附因子的R1R2区。扩增产物经双酶切后克隆到表达载体pET-32(a) 中。该重组质粒经酶切鉴定后,将其具有正确阅读框架的重组质粒转化到大肠杆菌BL21(DE3)感受态细胞,37℃下经IPTG诱导表达,得到相对分子质量约29000的融合蛋白,表达量约为11%。  相似文献   
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A bacterin containing serotypes 1 and 5 of Haemophilus pleuropneumoniae was developed for the prevention and the control of porcine pleuropneumonia. It was injected intramuscularly into three groups of ten piglets, the first group with one dose, the second one with two doses and the third one with three doses at two-week intervals. Another group of ten piglets did not receive the vaccine. All the piglets were then challenged by an aerosol of mixed suspensions of H. pleuropneumoniae serotypes 1 and 5. Two and three injections of vaccine completely prevented mortality, whereas half of the control piglets and of those receiving only one dose of vaccine died. All surviving piglets, both control and vaccinated, had severe signs of respiratory disease for at least 36 hours after exposure to challenge. Moreover, vaccination did not induce the production of antibodies at high titers. Local reactions were not noted after vaccination and at postmortem; ten weeks after the challenge, there were no signs of abscess formation or induration.  相似文献   
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During 1972 to 1974, 112 Escherichia coli strains isolated from diarrheic piglets were recieved from different parts of the Province of Quebec, Canada. Fifty-six strains elicited a positive gut loop response in three week old piglets and were then considered as Moon's class 1 enteropathogens, while four of the 56 remaining strains reacted only in ten day old piglets and were classified as class 2 enteropathogens. Forty-eight strains produced both a heat-labile and a heat-stable enterotoxin, while 12 isolates which included the four class 2 enteropathogens produced only a heat-stable enterotoxin. Fifty-one enterotoxigenic strains could be serogrouped using OK antisera against E. coli strains commonly associated with colibacillosis in piglets. The most common serogroups encountered were O157: K "V17"; 88a,c, O149:K91; 88a.c. O157:K"V1c, O149:K91; 88a.c, O157:K"V17"; 88 a,c or a,b and O45:K"E5"; 88a,c. No significant difference was observed in the fermentation patterns, antibiotic susceptibility, colicin production, production of a filterable hemolysin and transferable tetracycline resistance between the enterotoxigenic and the nonenterotoxigenic strains.  相似文献   
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A total of 20 (14 females, six males) captive striped skunks (Mephitis mephitis) with miniature temperature dataloggers implanted free in the abdominal cavity were examined for reproductive performance and pathology because of implants. Eleven of 12 female skunks reproduced successfully 45.9+/-3.7 days after surgery to remove implanted dataloggers. The pregnancy rate of 91.7% (11/12) was much higher than other captive studies and was comparable with that of wild skunks. Eight striped skunks (six males, two females) that were euthanatized and necropsied after having implants in the abdominal cavity for 5 mo showed no apparent pathology associated with the implant. Implantation of the intraperitoneal devices did not lead to complications in abdominal tissues. Neither implant nor surgery affected reproduction. We conclude that implanted dataloggers can safely be used in physiologic studies of striped skunks or possibly other small carnivores in captive or field studies.  相似文献   
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Black Aspergilli, and in particular Aspergillus carbonarius, are the main causes of contamination of grapes and their by-products by ochratoxin A. A PCR-based method was developed to detect DNA of A. carbonarius and A. japonicus. Two pairs of primers (CARBO1/2 and JAPO1/2) designed from the calmodulin gene, produced PCR products of 371 and 583 bp for A. carbonarius and A. japonicus, respectively. Primer specificity was tested with DNA of 107 strains belonging to Aspergillus section Nigri isolated mostly from grapes in Europe. The sensitivity of primers CARBO1/2 and JAPO1/2 was 12.5 pg when using pure total genomic DNA of the two species. The developed primers provide a powerful tool for detection of the main ochratoxigenic producing Aspergillus species in grapes.  相似文献   
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