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51.
Cloning and sequencing of the ovine gamma-interferon gene 总被引:1,自引:0,他引:1
Cytokines are major modulators of the immune system of all animals. The cloning and expression of recombinant cytokine genes have permitted the analysis of their immune function and role in the control of the immune response to disease and vaccination. While human, murine, and bovine genes have been cloned and sequenced, the cloning of ovine cytokine genes has not yet been reported. As sheep are of dominant economic importance to the Australian farming industry, it is of significance to clone and express these genes to facilitate the development of new and better vaccines and pharmaceuticals. We have initially selected ovine gamma-interferon (gamma-IFN) as a target cytokine gene. By the use of the polymerase chain reaction (PCR), using primers based on the bovine gamma-interferon sequence, we have amplified the ovine gamma-interferon gene from crude messenger RNA extracted from lymphocytes. After cloning and DNA sequencing the gene, we found that ovine gamma-IFN is 93% identical to bovine gamma-IFN in amino acid sequence. This result indicates that the PCR method will be a rapid and efficient means for cloning other ovine cytokine genes. 相似文献
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GA SMITH PL YOUNG† BJ RODWELL† MA KELLY† GJ STORIE† CA FARRAH† JS MATTICK† 《Australian veterinary journal》1994,71(3):65-70
SUMMARY An Australian bovine herpesvirus 1 (BHV1) isolate with a defined (427 base pair) deletion in the protein coding region of the thymidine kinase gene was obtained by standard marker rescue procedures. After selection in the presence of the nucleotide analogue 5iodo-deoxy-uridine the virus was analysed by hybridisation with three differential oligonucleotide probes, restriction endonuclease profile studies and DNA sequence analysis. The virus elicited an immune response in recipient animals after either intramuscular or intravenous administration and produced no significant deleterious side-effects when administered at a dose sufficient to stimulate the host immune response. The safety and immunogenicity of the recombinant BHV1 virus 39B1 were similar to those reported for other registered BHV1 vaccines and the virus would appear to be suitable for the production of a vaccine seed lot and more exhaustive field trials as a prelude to commercial vaccine production and registration. 相似文献
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Objective To determine the reliability of collecting serial 4-mL blood samples over 13 h using a 20-gauge, cephalic catheter in conscious dogs. Design Prospective, observational study. Animals Twelve (6 males, 6 females) healthy, neutered, lean dogs. Procedure Percutaneous placement of a 20-gauge, 1.1 × 30 mm intravenous over-the-needle catheter into the cephalic vein was performed in each dog on three occasions. Catheter patency was maintained by intermittent flushing with 0.9% saline and 1 IU/mL heparinised saline solutions. Blood samples (4 mL) were obtained at 10 time-points over 13 h, with close monitoring of the dogs and the indwelling catheters. Blood volume, resistance on aspirating blood sample (minimal or marked resistance) and the methods used to improve blood flow were recorded. Results A high proportion (93%) of blood sample collections achieved the required 4-mL volume, and the remaining samples were greater than 1.5 mL. Of the 358 collections via the cephalic catheter, 311 (87%) were obtained with ‘minimal resistance’. The remaining collections had ‘marked resistance’ (n=47) and were associated with a loose catheter in 11% (5/47) and of unknown cause in 89% (42/47). After ‘marked resistance’ had been encountered and the catheter was flushed with saline and heparin solutions, subsequent sampling with ‘minimal resistance’ was frequently possible from the same catheter. Conclusion Use of a percutaneous, 20-gauge intravenous cephalic catheter allowed reliable serial collection of 4-mL blood samples over 13 h in conscious dogs. 相似文献
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