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91.
Yen Kuan Ng Muhammad Ehsaan Sheryl Philip Mark M. Collery Clare Janoir Anne Collignon Stephen T. Cartman Nigel P. Minton 《PloS one》2013,8(2)
Sophisticated genetic tools to modify essential biological processes at the molecular level are pivotal in elucidating the molecular pathogenesis of Clostridium difficile, a major cause of healthcare associated disease. Here we have developed an efficient procedure for making precise alterations to the C. difficile genome by pyrE-based allelic exchange. The robustness and reliability of the method was demonstrated through the creation of in-frame deletions in three genes (spo0A, cwp84, and mtlD) in the non-epidemic strain 630Δerm and two genes (spo0A and cwp84) in the epidemic PCR Ribotype 027 strain, . The system is reliant on the initial creation of a pyrE deletion mutant, using Allele Coupled Exchange (ACE), that is auxotrophic for uracil and resistant to fluoroorotic acid (FOA). This enables the subsequent modification of target genes by allelic exchange using a heterologous pyrE allele from Clostridium sporogenes as a counter-/negative-selection marker in the presence of FOA. Following modification of the target gene, the strain created is rapidly returned to uracil prototrophy using ACE, allowing mutant phenotypes to be characterised in a PyrE proficient background. Crucially, wild-type copies of the inactivated gene may be introduced into the genome using ACE concomitant with correction of the pyrE allele. This allows complementation studies to be undertaken at an appropriate gene dosage, as opposed to the use of multicopy autonomous plasmids. The rapidity of the ‘correction’ method (5–7 days) makes pyrE
− strains attractive hosts for mutagenesis studies. R20291相似文献
92.
ZiMian Wang Stanley Heshka Kuan Zhang Carol N. Boozer Steven B. Heymsfield 《Obesity (Silver Spring, Md.)》2001,9(5):331-336
There are many published methods for predicting resting energy expenditure (REE) from measured body composition. Although these published reports extend back almost a century, new related studies appear on a regular basis. It remains unclear what the similarities and differences are among these various methods and what, if any, advantages the newly introduced REE prediction models offer. These issues led us to develop an organizational system for REE prediction methods with the goal of clarifying prevailing ambiguities in the field. Our classification scheme is founded on body composition level (whole‐body, tissue‐organ, cellular, and molecular) and related components as the REE predictor variables. Each existing REE prediction method by body composition must belong to one body composition level. The suggested classification system, founded on a conceptual basis, highlights similarities and differences among the diverse REE‐body composition prediction methods, provides a framework for teaching REE‐body composition relationships, and identifies important future research opportunities. 相似文献
93.
植物抗细菌病害基因工程研究进展和展望 总被引:1,自引:0,他引:1
综述了利用基因工程提高植物对细菌病害抗性的各种方法,包括利用非植物抗菌蛋白,抑制细菌的致病或毒性因子,增强植物本身的抗病能力和人工诱导侵染点细胞程序化坏死。这些方法的成功都与抗菌化合物的作用机制及植物和病原细菌之间的相互作用的分子生物学的研究密切相关,还展望了这些方法的应用前景。 相似文献
94.
Jasmonates are a new class of plant hormones that play important roles in plant development and plant defense. TheCOI1 gene was previously shown to be required for jasmonate-regulated plant fertility and defense. We demonstrated for the first
time that COI1 interacts with theArabidopsis SKP1-LIKE1 (ASK1) to form a complex that is required for jasmonate action inplanta. Functional analysis by antisense strategy showed thatASK1 is involved in male fertility. 相似文献
95.
Jasmonates are a new class of plant hormones that play important roles in plant development and plant defense. The COI1 gene was previously shown to be required for jasmonate-regulated plant fertility and defense. We demonstrated for the first time that COI1 interacts with the Arabidopsis SKP1-LIKE1 (ASK1) to form a complex that is required for jasmonate action in planta. Functional analysis by antisense strategy showed that ASK1 is involved in male fertility. 相似文献
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Alpha-Chymotrypsin was light sensitized by acylating with cis-cinnamoyl ester, a substrate interconvertible to the trans form by ultraviolet (UV) light. The degree of acylation by this method was complete leaving no residual activity of the enzyme. Upon UV irradiation the inhibited enzyme regained about 70% of its original activity, thereby adding light-sensitiveness to the proteolytic enzyme. In seeking a photographic application of the light-sensitized enzyme, a pigmenting enzyme was incorporated with it. The coupled enzyme system was shown to exhibit a light signal in the form of dark pigment slurry. 相似文献
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