全文获取类型
收费全文 | 1038篇 |
免费 | 61篇 |
国内免费 | 51篇 |
专业分类
1150篇 |
出版年
2023年 | 8篇 |
2022年 | 7篇 |
2021年 | 25篇 |
2020年 | 11篇 |
2019年 | 25篇 |
2018年 | 22篇 |
2017年 | 27篇 |
2016年 | 15篇 |
2015年 | 29篇 |
2014年 | 38篇 |
2013年 | 43篇 |
2012年 | 32篇 |
2011年 | 42篇 |
2010年 | 33篇 |
2009年 | 44篇 |
2008年 | 48篇 |
2007年 | 54篇 |
2006年 | 58篇 |
2005年 | 50篇 |
2004年 | 50篇 |
2003年 | 38篇 |
2002年 | 58篇 |
2001年 | 42篇 |
2000年 | 36篇 |
1999年 | 23篇 |
1998年 | 11篇 |
1997年 | 17篇 |
1996年 | 22篇 |
1995年 | 26篇 |
1994年 | 19篇 |
1993年 | 19篇 |
1992年 | 19篇 |
1991年 | 15篇 |
1990年 | 16篇 |
1989年 | 9篇 |
1988年 | 15篇 |
1987年 | 15篇 |
1986年 | 6篇 |
1985年 | 18篇 |
1984年 | 19篇 |
1983年 | 8篇 |
1982年 | 16篇 |
1981年 | 9篇 |
1980年 | 1篇 |
1979年 | 6篇 |
1978年 | 1篇 |
1976年 | 1篇 |
1975年 | 1篇 |
1972年 | 1篇 |
1971年 | 1篇 |
排序方式: 共有1150条查询结果,搜索用时 9 毫秒
21.
The human multiple drug resistance (MDR) gene has been used as a model for human gene transfer which could lead to human gene therapy. MDR is a transmembrane protein which pumps a number of toxic substances out of cells including several drugs used in cancer chemotherapy. Normal bone marrow cells express low levels of MDR and are particularly sensitive to the toxic effects of these drugs. There are two general applications of MDR gene therapy: (1) to provide drug-resistance to the marrow of cancer patients receiving chemotherapy, and (2) as a selectable marker which when co-transferred with a non-selectable gene such as the human beta globin gene can be used to enrich the marrow for cells containing both genes. We demonstrate efficient transfer and expression of the human MDR gene in a retroviral vector into live mice and human marrow cells including CD34+ cells isolated from marrow and containing the bulk of human hematopoietic progenitors. MDR gene transduction corrects the sensitivity of CD34+ cells to taxol, an MDR drug substrate, and enriches the marrow for MDR-transduced cells. The MDR gene-containing retroviral supernatant used has been shown to be safe and free of replication-competent retrovirus. Because of the safety of the MDR retroviral supernatant, and efficient gene transfer into mouse and human marrow cells, a phase 1 clinical protocol for MDR gene transfer into cancer patients has been approved to evaluate MDR gene transfer and expression in human marrow. 相似文献
22.
综述了近年来有关利用基因转移技术修饰肿瘤细胞制备肿瘤基因工程疫苗的最新研究进展,着重阐述了逆转录病毒载体介导的基因转移及其安全性;归纳了目前可用于肿瘤基因工程疫苗的各种目的基因的特点及作用并对这类肿瘤疫苗制备过程中所存在的问题进行了分析. 相似文献
23.
Transformation of Kluyveromyces lactis with the kanamycin (G418) resistance gene of Tn903 总被引:11,自引:0,他引:11
Direct selection of Kluyveromyces lactis resistant to the antibiotic G418 following transformation with the kanamycin resistance gene of Tn903 required the development of a procedure for producing high yields of viable spheroplasts and for the isolation of autonomous replication sequences (ARS). To obtain high yields of viable spheroplasts, cells were treated with (1) a thiol-reducing agent (L-cysteine), and (2) a high concentration of an osmotic stabilizer, 1.5 M sorbitol. Several ARS-containing plasmids were selected from a K. lactis recombinant DNA library in K. lactis and in Saccharomyces cerevisiae. Two of four ARS clones selected in K. lactis promoted transformation frequencies of 5-10 X 10(2) G418-resistant cells/micrograms of plasmid DNA. This frequency of transformation was at least twice as high as with ARS clones selected in S. cerevisiae. The stability of ARS-containing plasmids varied; after 20 generations of growth in the presence of G418, 16-38% of the cells remained resistant to the drug. In the absence of selection pressure less than 5% of the cells retained the drug-resistance phenotype. Plasmids containing the ARS1 or 2 mu replicon of S. cerevisiae failed to transform K. lactis for G418 resistance. Inclusion of S. cerevisiae centromere, CEN4, in a K. lactis ARS recombinant plasmid did not increase the stability of the plasmid in K. lactis, and marker genes on the vector segregated predominantly 4-:0+ through meiosis. We conclude that neither the ARS sequences or the centromere of S. cerevisiae was functioning in K. lactis. 相似文献
24.
25.
We examined the survival of a host Escherichia coli K-12 bacterium containing two transferable plasmids (pLM2, pSL222-4) and one poorly mobilizable plasmid (pBR322), and the transfer of these three plasmids to endogenous bacteria in the human intestinal tract. The survival of this plasmid-carrying host organism in four human volunteers was 3.5 to 6 days at recovery rates of 10?1 to 10?4. This finding was similar to our previous survival data on the same organism bearing a single plasmid. The K-12 strain appeared to be under a strong selective disadvantage in the human gut, since, even when bearing a tetracycline-resistant plasmid, its titer did not increase despite the administration of tetracycline. Studies of transferability showed that, while the transfer-depressed incFII plasmid pSL222-4 transferred at a frequency of 10?1 in culture, its transfer in the human gut was much less frequent. The number of new recipients per donor cell ingested was about 10?5, which included new recipients arising by multiplication. The recovery of pSL222-4 transcipients was enhanced by the administration of tetracycline on day 6. Neither the transfer-repressed, broad host range incP plasmid pLM2, nor the plasmid pBR322, could be detected in any endogenous host bacteria. Using the transfer and mobilization frequencies obtained in culture and the number of new recipients of pSL222-4 in the intestinal tract, we estimated that any in vivo mobilization of pBR322 to a new recipient could not occur at a frequency higher than 10?12. 相似文献
26.
Use of lambda phasmids for deletion mapping of non-selectable markers cloned in plasmids 总被引:5,自引:0,他引:5
A nonselectable gene carried on a poorly selectable recombinant plasmid has been physically mapped by deletion analysis. Our method involved cloning the plasmid into a coliphage lambda vector and treating the recombinant phage with a chelator. Virtually all particles surviving this treatment carried large deletions within the plasmid insert. Further deletion analysis was done by inserting a selectable lambda sequence into one such deletion derivative and repeating the chelator selection. Chelator selection was also used to isolate deletions constructed in vitro. The deleted phage are readily characterized by restriction mapping, and the gene in question scored after infection of a mutant host strain. These techniques have enabled us to physically assign the cyclopropane fatty acid synthase gene of Escherichia coli to 0.8 kb of a 16-kb segment after characterizing only a small number of isolates. This approach should be generally useful in the mapping of plasmids for which no convenient method exists for selecting or scoring the gene in question. 相似文献
27.
N. C. Smith 《International journal for parasitology》1992,22(8):1047-1082
28.
29.
30.