全文获取类型
收费全文 | 744篇 |
免费 | 25篇 |
国内免费 | 16篇 |
专业分类
耳鼻咽喉 | 4篇 |
妇产科学 | 2篇 |
基础医学 | 167篇 |
口腔科学 | 11篇 |
临床医学 | 80篇 |
内科学 | 71篇 |
皮肤病学 | 16篇 |
神经病学 | 7篇 |
特种医学 | 4篇 |
外科学 | 44篇 |
综合类 | 120篇 |
预防医学 | 129篇 |
眼科学 | 2篇 |
药学 | 115篇 |
中国医学 | 2篇 |
肿瘤学 | 11篇 |
出版年
2024年 | 1篇 |
2023年 | 1篇 |
2022年 | 3篇 |
2021年 | 8篇 |
2020年 | 5篇 |
2019年 | 14篇 |
2018年 | 8篇 |
2017年 | 5篇 |
2016年 | 10篇 |
2015年 | 10篇 |
2014年 | 34篇 |
2013年 | 42篇 |
2012年 | 37篇 |
2011年 | 44篇 |
2010年 | 39篇 |
2009年 | 38篇 |
2008年 | 37篇 |
2007年 | 55篇 |
2006年 | 37篇 |
2005年 | 42篇 |
2004年 | 32篇 |
2003年 | 23篇 |
2002年 | 20篇 |
2001年 | 18篇 |
2000年 | 15篇 |
1999年 | 18篇 |
1998年 | 23篇 |
1997年 | 16篇 |
1996年 | 14篇 |
1995年 | 19篇 |
1994年 | 23篇 |
1993年 | 16篇 |
1992年 | 17篇 |
1991年 | 6篇 |
1990年 | 17篇 |
1989年 | 3篇 |
1988年 | 12篇 |
1987年 | 7篇 |
1986年 | 1篇 |
1985年 | 8篇 |
1983年 | 1篇 |
1982年 | 3篇 |
1980年 | 1篇 |
1978年 | 1篇 |
1977年 | 1篇 |
排序方式: 共有785条查询结果,搜索用时 15 毫秒
1.
2.
本文以 E.coli CM891为靶细胞,用细菌内抗突变作用模式研究了肉桂醛,鞣酸,二烯丙三硫的抗4NQO 突变性及其作用机制。含质粒 pKM101的 CM891的高抗株(抗50μg/ml 氨苄青霉素)能提高该菌株的自发突变率和4NQO 诱发的突变率以及对鞣酸的杀伤抗性。肉桂醛的抗突变性不依赖质粒 pKM101效应,但与暂时性生长延搁有关。鞣酸及二烯丙三硫的抗突变机制可能包括质粒 pKM101介导的易误修复。上述三种化学物中每二种联合应用均显示抗4NQO 突变性的协同效应及对靶细胞的毒性杀伤作用。 相似文献
3.
赖型钩体flaB2与VR1012中的CpG基序分析 总被引:3,自引:0,他引:3
目的:对问号赖型钩端螺旋体(赖型钩体)DNA疫苗[包括内鞭毛蛋白基因(flaB2)和质粒DNA表达载体(VR1012)]的CpG基序(CpG motifs)进行分析,为DNA疫苗免疫机制的阐明和提高DNA疫苗的效能奠定基础。方法:以flaB2与VR1012构建重组DNA的免疫原,对flaB2及VR1012全核苷酸序列进行计算机分析(分类、计数和定位)。结果:CpG的“C”的侧翼为两个嘌呤,“G”的侧翼为两个嘧啶,在flaB2中共3个,分别为GACGCT,GACGTC和GACGCC;在VR1012中共11个,分别为GACGTC1个,GACGCT2个,GACGCC1个,GACGTT1个,GGCGTT2个,GGCGCT2个,GGCGCC1个,AACGCT1个,其中特别重要的TGACGTCA4个和TAACGCCA有1个,位于5'端456-463;509-516;592-599;778-785和486-493;4个TGACGTCA和1个TAACGCCA均位于5'端且相对集中。结论:赖型钩体flaB2与VR1012构成的DNA疫苗含有TGACGTCA等CpG,这些基序又称免疫刺激序列,构成了DNA疫苗中的佐剂。 相似文献
4.
结核分枝杆菌Ag85B/GST融合蛋白表达载体构建及在大肠杆菌中的表达 总被引:1,自引:1,他引:0
目的 构建结核分枝杆菌Ag85B/GSF融合蛋白表达质粒,并在大肠杆菌(E.coli)中诱导表达。方法 以质粒pUC18/Ag85B为模板,用PCR法扩增结核杆菌Ag85B基因,扩增的Ag85B上游含有EcoR Ⅰ酶切位点,下游含有SalⅠ酶切位点;将Ag85B基因定向插入质粒pGEX-4T-1中,构建原核表达质粒pGEX-Ag85B并转化E.coli B121,筛选阳性重组子,限制性内切酶切鉴定和DNA序列测定,异丙基硫代半乳糖苷(IPTG)诱导表达,SDS-PAGE和Westem blot鉴定结果。结果 成功构建原核表达质粒pGEX-Ag85B并表达出Mr约56000大小的Ag85B/GSY融合蛋白,表达量占总菌体的30%。结论 为进一步进行纯化Ag85B蛋白和研究其在膀胱肿瘤治疗中的作用奠定了基础。 相似文献
5.
新生儿病房铜绿假单胞菌的血清学分型,质粒分析研究 总被引:1,自引:0,他引:1
新生儿病房1989年10月~1990年12月、1996年3~6月两个时间段分离40株铜绿假单胞菌,血清学分型率97.5%,第一时间段以O11型最多,占30.4%,集中在1990年6~10月,第二时间段O3型占93.8%,集中在1996年3~6月,表明该病房发生两次铜绿假单胞菌医院感染流行。质粒检出率42.9%,O11型菌株质粒谱为8.1、46.5、179.1kb,并经HindⅢ酶切进一步证实为同一克隆。对氟哌酸、阿米卡星及头孢他啶的敏感率分别为100%、97.3%和95%。对庆大霉素耐药率明显上升,提示控制铜绿假单胞菌医院感染的重要性。 相似文献
6.
Francisco Fierro Katarina Kosalková Santiago Gutiérrez Juan F. Martín 《Current genetics》1996,29(5):482-489
Plasmid vectors containing theAMA1 sequence transformed with high efficiency and replicated autonomously inPenicillium chrysogenum. The efficiency of transformation ofP. chrysogenum was related to the length of theAMA1 fragment used for constructing the different autonomously replicating plasmids. One of the two palindromic inverted repeats ofAMA1 (the 2.2-kbSalI-HindIII fragment) is sufficient to confer autonomous replication and a high transformation efficiency. Deletion of the 0.6-kb central fragment located between the inverted repeats did not affect either the ability of the plasmids to replicate autonomously or the efficiency of transformation, but did alter the mitotic stability and the plasmid copy number. Deletion of any fragment of the 2.2-kb repeat caused the loss of the ability to replicate autonomously and reduced the transformation efficiency. Most of the transformants retained the original plasmid configuration, as multimers and without reorganization, after several rounds of autonomous replication. TheAMA1 region works as an origin of replication inP. chrysogenum andA. nidulans but not apparently inAcremonium chrysogenum. 相似文献
7.
Summary We have studied the structure of the two linear DNA plasmids, kl and k2, present in killer strains of Kluyveromyces lactis. Two killer strains of different origins, CBS 2359 and IFO 1267 were examined. For both strains, identical restriction maps of kl and k2 DNA were obtained. Several restriction sites previously reported for the kl DNA of the strain IFO 1267 have been confirmed. The molecular weights of these double-stranded DNAs were 8.8 kilobase pairs for kl and 13.4 for k2, as determined by electrophoresis of restriction fragments. The plasmid DNA from a nonkiller mutant, NK2/1, was also examined. In this mutant, the kl DNA was replaced by a smaller DNA (5.9 kilobase pairs), the k2 DNA being normal. Restriction enzyme analysis showed that the new plasmid DNA was also linear. Hybridization experiments demonstrated that it was derived from the kl DNA by deletion of a 2.9 kilobase pair segment from the central part of the kl DNA. The deleted segment carries a gene involved in toxin production, but is not related to immunity since the mutant is resistant to killers. The plasmid DNA of K. lactis showed no detectable sequence homology with the double stranded RNA of the killer system of Saccharomyces cerevisiae. Neither was any homology found with nuclear and mitochondria) DNA. 相似文献
8.
《Research in microbiology》2021,172(6):103870
We previously reported the complete genome of Streptomyces lavendulae subsp. lavendulae CCM 3239, containing the linear chromosome and the large linear plasmid pSA3239. Although the chromosome exhibited replication features characteristic for the archetypal end-patching replication, it lacked the tap/tpg gene pair for two proteins essential for this process. However, this archetypal tpgSa-tapSa operon is present in pSA3239. Complete genomic sequence of the S. lavendulae Del-LP strain lacking this plasmid revealed the circularization of its chromosome with a large deletion of both arms. These results suggest an essential role of pSA3239-encoded TapSa/TpgSa in the end-patching replication of the chromosome. 相似文献
9.
华南地区质粒介导超广谱β-内酰胺酶的基因分型研究 总被引:59,自引:4,他引:59
目的:了解华南地区质粒介导超广谱β-内酰胺酶(ESBLs)的发生率及基因型特征。方法:收集2001年4月-9月革兰阴性菌临床分离无重复株共1184株,采用NCCLS表型筛选和确认试验进行了ESBLs产酶的识别,E-test法检测各亚型ESBLs的MICs值,质粒接合及电转化实验,耐药质粒提取及酶切指纹分析,等电聚焦电泳,PCR通用引物扩增TEM、SHV、CTX-M、VEB、PER、SFO基因及其克隆测序进行ESBLs基因分型和质粒定位。结果:革兰阴性苗ESBLs的检出率为14.6%(173/1184);获得产ESBLs接合子67株,电转化子11株,其中产CTX-M-14型ESBLs为33.3%(26/78)、CTX-M-3为23.1%(18/78)、CTX-M-9为14.1%(11/78)及SHV-2a为2.6%(2/78),未定型为5.1%(4/78);29.5%(23/78)野生株伴广谱酶TEM-1或SHV-1型;各型ESBLs基因约定位在35-190kb大小的可接合性低执行者拷贝数天然质粒上;CTX-M型ESBLs以对头孢噻肟高水平高耐为特征。结论:华南地区质粒介导的ESBLs以CTX-M型衍生酶为主,其次是SHV型酶。 相似文献
10.
Luby TM Cole G Baker L Kornher JS Ramstedt U Hedley ML 《Clinical immunology (Orlando, Fla.)》2004,112(1):45-53
Injection of microparticle-encapsulated DNA elicits immune responses to plasmid-encoded antigens in mice and humans. Cytochrome P450 CYP1B1 (CYP1B1) is a member of the CYP1 P450 enzyme family that is overexpressed in a variety of solid tumors. The work described herein was performed to study the kinetics of stimulating T cell responsiveness with an encapsulated DNA encoding CYP1B1 and provides support for the clinical development of this formulation. Immunization of HLA-A2/Kb transgenic mice with human CYP1B1 encoding plasmid DNA formulated in poly(lactide-co-glycolide) (PLG) microparticles elicits CD8+ T cells that respond to human CYP1B1-positive target cells. The duration of the immune response, the effect on the immune response of multiple injections, and the safety of repeated injections were studied. These results show that the PLG-encapsulated DNA therapeutic elicits durable immune responses to CYP1B1, the responses are dependent on repeat immunization, and that the formulation is well tolerated. 相似文献