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171.
pepT基因编码一种金属依赖性肽酶T (peptidase T,PepT),能特异性催化三肽N端氨基酸,因此也称为氨肽酶T。研究发现大多数氨肽酶参与细菌蛋白质新陈代谢和调节三肽活性,但关于PepT在细菌毒力及致病性方面的报道较少。[目的]本文选取PepT为研究对象,研究其对副溶血弧菌生物学特性及致病性的影响。[方法]通过构建缺失株ΔpepT和回补株CΔpepT,比较菌株在运动性、生物被膜、环境耐受、细胞毒性等方面的差异。[结果]与野生株相比,ΔpepT缺失株的极性鞭毛转录水平极显著下降,浮游运动能力降低;同时生物被膜形成能力减弱,而细菌群集运动及环境耐受能力无显著差异。此外,缺失pepT基因会导致副溶血弧菌的细胞毒性和小鼠毒力作用显著下降。[结论]pepT基因与副溶血弧菌浮游运动和生物被膜形成能力相关,并且影响其致病性。 相似文献
172.
Analysis of Vibrio seventh pandemic island II and novel genomic islands in relation to attachment sequences among a wide variety of Vibrio cholerae strains 下载免费PDF全文
Tuan Hai Nguyen Tho Duc Pham Naomi Higa Hanako Iwashita Taichiro Takemura Makoto Ohnishi Kouichi Morita Tetsu Yamashiro 《Microbiology and immunology》2018,62(3):150-157
173.
A pure culture of the luminous bacterium Vibrio fischeri is maintained in the light-emitting organ of the sepiolid squid Euprymna scolopes. When the juvenile squid emerges from its egg it is symbiont-free and, because bioluminescence is part of an anti-predatory behavior, therefore must obtain a bacterial inoculum from the surrounding environment. We document here the kinetics of the process by which newly hatched juvenile squids become infected by symbiosis-competent V. fischeri. When placed in seawater containing as few as 240 colony-forming-units (CFU) per ml, the juvenile became detectably bioluminescent within a few hours. Colonization of the nascent light organ was initiated with as few as 1 to 10 bacteria, which rapidly began to grow at an exponential rate until they reached a population size of approximately 105 cells by 12 h after the initial infection. Subsequently, the number of bacteria in the established symbiosis was maintained essentially constant by a combination of both a >20-fold reduction in bacterial growth rate, and an expulsion of excess bacteria into the surrounding seawater. While V. fischeri cells are normally flagellated and motile, these bacteria did not elaborate these appendages once the symbiosis was established; however, they quickly began to synthesize flagella when they were removed from the light organ environment. Thus, two important biological characteristics, growth rate and flagellation, were modulated during establishment of the association, perhaps as part of a coordinated series of symbiotic responses. 相似文献
174.
A method for studying the biotype El Tor associated mannose-sensitive haemagglutinin (MSHA) of V. cholerae O1 has been developed. By using crude MSHA adsorbed to chicken erythrocytes as solid phase antigen in an enzyme-linked immunosorbent assay (ELISA), antisera against V. cholerae of the El Tor biotype reacted in high titre with the MSHA-coated cells, whereas antisera against vibrios of the classical biotype did not bind significantly, i.e. in higher titre than pre-immune sera. The binding of anti-MSHA serum, or a monoclonal antibody against MSHA, to the MSHA-coated erythrocytes could be efficiently inhibited by crude MSHA as well as by El Tor vibrios whereas neither V. cholerae lipopolysaccharide nor different strains of classical vibrios had any inhibitory effect. These results support the existence of an El Tor-associated immunogen. They also suggest a possibility of determining antibodies against different haemagglutinins in ELISA without having access to purified antigens. 相似文献
175.
对襄阳市中心医院分离的2株疑似霍乱弧菌进行鉴定及药物敏感性试验。利用MicroScan WalkAway 40鉴定仪进行生化鉴定及药物敏感性试验,玻片凝集法确定血清型别,PCR扩增16SrRNA保守区基因并将产物进行测序分析。此2株疑似霍乱菌株O1群及O139群霍乱弧菌诊断血清均不凝集,16SrRNA扩增产物测序Blast比对分析与数据库中霍乱弧菌相似性达100%,药敏结果显示对氨苄西林、庆大霉素、环丙沙星、阿米卡星、氯霉素、复方新诺明(SXT)、四环素均敏感。该病例为非O1、非O139群霍乱弧菌导致的败血症,可能经胃肠道途径传播。 相似文献
176.
Mathew A. Maliakal Mepur H. Ravindranath Reiko F. Irie Donald L. Morton 《Glycoconjugate journal》1994,11(2):97-104
In the measurement of total lipid-bound sialic acids involving periodic acid oxidation, as in the periodate-resorcinol assay, the inner sialic acids of disialoglycolipids (such as GD3 and GD2) are not involved because their 2,8 ketosidic linkages are resistant to periodic acid oxidation, even after acid/enzyme hydrolysis or alkali pretreatment. However, the sialic acids from these glycolipids can be recovered completely after cleavage of 2,8 linkages byV. cholerae sialidase in the presence of cholic acid, sodium dodecyl sulphate and calcium. Interestingly, removal of calcium or detergent(s) or both significantly minimizes the sialidase action on the disialyl residues of these gangliosides. Therefore, we recommend sialidase (Vibrio cholerae) pretreatment of the glycolipids in the presence of cholic acid, SDS and Ca2+ for complete recovery of sialic acids from di- and polysialogangliosides and for accurate measurement of total lipid-bound sialic acids by periodate-resorcinol assay.Presented at the Second International Glycobiology Symposium which was held in San Francisco, CA, USA (14 February 1994). 相似文献
177.
toxR基因作为荧光定量PCR靶基因设计TaqMan探针快速检测副溶血弧菌 总被引:12,自引:0,他引:12
以toxR基因为靶基因,通过优化反应条件建立了快速检测副溶血弧茵的TaqMan实时荧光PCR方法.特异性试验表明,该方法能选择性检测副溶血弧茵,而与金黄色葡萄球菌、沙门氏菌、单增李斯特杆菌等多种常见的食源性病原茵没有交叉反应:灵敏度试验表明,该方法最少可检测到25个拷贝的toxR基因重组质粒,对纯培养物和模拟食品样品直接检测的灵敏度分别为21 cfu/mL和210 cfu/g;重复性试验表明,同一样品于试验内及试验间的变异系数分别为0.9%和1.3%:所制作的标准曲线在2.5 × 101~2.5 × 106拷贝数之间有较好的线性关系,能对副溶血孤菌进行准确的定量分析.结果表明,本研究所建立的副溶血弧菌实时荧光PCR检测方法具有特异性好,灵敏度高、重复性好的特点,能进行定量检测,而且检测时间从核酸抽提到出实验结果仅需要3 h.是快速检测副溶血弧菌的有效手段. 相似文献
178.
Sreerupa Ganguly Amarshi Mukherjee Budhaditya Mazumdar Amar N. Ghosh Kalyan K. Banerjee 《The Journal of biological chemistry》2014,289(7):4001-4008
Vibrio cholerae cytolysin/hemolysin (VCC) is an amphipathic 65-kDa β-pore-forming toxin with a C-terminal β-prism lectin domain. Because deletion or point mutation of the lectin domain seriously compromises hemolytic activity, it is thought that carbohydrate-dependent interactions play a critical role in membrane targeting of VCC. To delineate the contributions of the cytolysin and lectin domains in pore formation, we used wild-type VCC, 50-kDa VCC (VCC50) without the lectin domain, and mutant VCCD617A with no carbohydrate-binding activity. VCC and its two variants with no carbohydrate-binding activity moved to the erythrocyte stroma with apparent association constants on the order of 107
m−1. However, loss of the lectin domain severely reduced the efficiency of self-association of the VCC monomer with the β-barrel heptamer in the synthetic lipid bilayer from ∼83 to 27%. Notably, inactivation of the carbohydrate-binding activity by the D617A mutation marginally reduced oligomerization to ∼77%. Oligomerization of VCC50 was temperature-insensitive; by contrast, VCC self-assembly increased with increasing temperature, suggesting that the process is driven by entropy and opposed by enthalpy. Asialofetuin, the β1-galactosyl-terminated glycoprotein inhibitor of VCC-induced hemolysis, promoted oligomerization of 65-kDa VCC to a species that resembled the membrane-inserted heptamer in stoichiometry and morphology but had reduced global amphipathicity. In conclusion, we propose (i) that the β-prism lectin domain facilitated toxin assembly by producing entropy during relocation in the heptamer and (ii) that glycoconjugates inhibited VCC by promoting its assembly to a water-soluble, less amphipathic oligomer variant with reduced ability to penetrate the bilayer. 相似文献
179.
180.
G.D. Ramirez G.W. Buck A.K. Smith K.V. Gordon J.B. Mott 《Journal of applied microbiology》2009,107(6):2047-2053
Aims: To determine the occurrence of the human pathogen, Vibrio vulnificus, in south Texas coastal waters. Methods and Results: Coastal waters were sampled monthly between August 2006 and July 2007. Water temperature, dissolved oxygen, pH, salinity, conductivity and turbidity were measured during each sampling event. Culture‐based techniques utilizing Vibrio vulnificus agar (VVA) and membrane‐Enterococcus indoxyl‐β‐d ‐glucoside agar (mEI) were used to assess the occurrence and levels of V. vulnificus and the faecal contamination indicator group, enterococci, respectively. Vibrio vulnificus isolates were confirmed using colony‐blot hybridization with the species‐specific VVAP probe. Vibrio vulnificus was isolated at all sites throughout the year even when the water temperature dropped to 9·71°C. Significant correlations were found between concentrations of V. vulnificus and the abiotic factors, water temperature (P = 0·002) and dissolved oxygen (P = 0·028), as well as between concentrations of V. vulnificus and enterococci (P < 0·001). Conclusions: This study demonstrated the year‐round presence of V. vulnificus in coastal waters of south Texas. Significance and Impact of the Study: These findings indicate that the potential for human exposure to the pathogen, V. vulnificus, exists throughout the year. It also suggests that routinely monitored data might be used to predict the occurrence of the pathogen. 相似文献