全文获取类型
收费全文 | 53257篇 |
免费 | 3798篇 |
国内免费 | 23篇 |
专业分类
57078篇 |
出版年
2024年 | 59篇 |
2023年 | 198篇 |
2022年 | 632篇 |
2021年 | 1014篇 |
2020年 | 627篇 |
2019年 | 772篇 |
2018年 | 1163篇 |
2017年 | 1012篇 |
2016年 | 1658篇 |
2015年 | 2558篇 |
2014年 | 2982篇 |
2013年 | 3329篇 |
2012年 | 4436篇 |
2011年 | 4244篇 |
2010年 | 2689篇 |
2009年 | 2427篇 |
2008年 | 3384篇 |
2007年 | 3228篇 |
2006年 | 2831篇 |
2005年 | 2638篇 |
2004年 | 2455篇 |
2003年 | 2100篇 |
2002年 | 1813篇 |
2001年 | 1485篇 |
2000年 | 1405篇 |
1999年 | 1124篇 |
1998年 | 453篇 |
1997年 | 384篇 |
1996年 | 273篇 |
1995年 | 245篇 |
1994年 | 237篇 |
1993年 | 196篇 |
1992年 | 387篇 |
1991年 | 339篇 |
1990年 | 304篇 |
1989年 | 268篇 |
1988年 | 200篇 |
1987年 | 190篇 |
1986年 | 152篇 |
1985年 | 126篇 |
1984年 | 91篇 |
1983年 | 98篇 |
1982年 | 75篇 |
1981年 | 65篇 |
1980年 | 65篇 |
1979年 | 78篇 |
1978年 | 66篇 |
1977年 | 55篇 |
1975年 | 55篇 |
1974年 | 74篇 |
排序方式: 共有10000条查询结果,搜索用时 0 毫秒
21.
22.
Secretion of Recombinant Pediocin PA-1 by Bifidobacterium longum, Using the Signal Sequence for Bifidobacterial α-Amylase 下载免费PDF全文
Gi-Seong Moon Yu-Ryang Pyun Myeong Soo Park Geun Eog Ji Wang June Kim 《Applied microbiology》2005,71(9):5630-5632
A recombinant DNA, encoding the chimeric protein of the signal sequence for bifidobacterial α-amylase mature pediocin PA-1, was introduced into Bifidobacterium longum MG1. Biologically active pediocin PA-1 was successfully secreted from the strain and showed bactericidal activity against Listeria monocytogenes and the same molecular mass as native pediocin PA-1. 相似文献
23.
24.
25.
26.
88 rice and 75 soybean samples were collected from 8 provinces of Korea from March through September in 1988. The Fusarium mycotoxins, zearalenone was analyzed by direct competitive enzyme linked immunosorbent assay. 10.2% of rice and 9.3 % of soybean samples contained detectable zearalenone. The average levels of zearalenone of rice and soybean samples were 11.78μg/kg and 7.70μ/kg, respectively. 相似文献
27.
To investigate the mechanism of inclusion body formation and the effect of a hydrophobic sequence on the in vivo polypeptide folding, the aggregation caused by recombinant fusion beta-galactosidase in Escherichia coli was examined. Two plasmids were constructed: pTBG(H-) carried only the preS2 sequence of the hepatitis B virus surface antigen (HBsAg) in front of the beta-galactosidase gene (lacZ) while pTBG(H+) carried an additional sequence encoding the amino-terminal hydrophobic sequence of the S region of HBsAg between preS2 and lacZ. Unlike cells expressing the fusion protein not containing the hydrophobic sequence, E. coli JM109/pTBG(H+) exhibited temperature-sensitive production of beta-galactosidase. As the culture temperature increased the activity decreased dramatically. This decrease in activity was not due to a decrease in fusion polypeptide production, but rather the fusion polypeptides containing the hydrophobic sequence aggregated within the cells at high temperature. However once the fusion polypeptides folded into proper conformation at low temperature, they maintained the activity even at high temperature. The results indicate that aggregation is a consequence of incorrect folding and assembly of the polypeptides, and is not derived from the native structure. The aggregates of the pTBG(H+)-encoded fusion polypeptides did not revert to active form when the culture temperature was lowered. 相似文献
28.
Yarrowia lipolytica KCCM50506, which transforms isobutyric acid to L-#-hydroxy isobutyric acid (L-#-HIBA), was screened. Chemostat cultures were carried out in jar fermentors at dilution rates of 0.02 hу to 0.12 hу. L-#-HIBA fermentation-regulating factors were determined to be specific growth rate, and concentrations of glucose and isobutyric acid in fermentor from analysis of steady-state data. The specific productivity of L-#-HIBA increased as the specific growth rate increased, apparently as a growth-associated type of product formation. A fed-batch culture was carried out under optimum conditions where the concentrations of glucose and isobutyric acid in the fermentor were maintained at 23 g lу and 9 g lу, respectively. The concentrations of cells and L-#-HIBA obtained at the end of fermentation were 20 g lу and 49 g lу, respectively, corresponding to 2.0 and 2.7 times more than concentrations in batch culture. 相似文献
29.
Sung Ho Son Sung Mee Choi Kum Boo Choi Yun Hee Lee Dea Sook Lee Myung Suk Choi Young Goo Park 《Biotechnology and Bioprocess Engineering》1999,4(2):112-118
Calli were induced from 300,000 embryos isolated from immature to mature stage of seeds collected on late September from 14
elite trees. When the embryos were cultured onto plastic Petri-dish containing 20 mL of modified B5 basal medium supplemented
with 3% (w/v) sucrose, 500 mg/L casein hydrolysate, 250 mg/L myo-inositol, 0.5% (w/v) polyvinyl polypyrrolidon (PVPP), 2×MS
vitamins, 0.5 mg/L gibberellic acid, and 10 mg/L 2,4-D after 2 weeks of culture, yellowish-white calli were immediately formed
on the surfaces of embryos, and subcultured for 4 weeks in same culture medium. Because most of calli maintained for more
than 3 months were revealed differences in their colors, surface texture, and growth rate, visual selection was made for first
round screening. When the size of visually selected calli larger than 19 mm in their diameter were inoculated, persistent
proliferation was observed. Among the plating methods tested for the selection of rapid growing cell lines at single cell
and/or small cell aggregate level, 2-layer spread plating revealed as the best for single cell cloning. To enhance cell growth
and maintain high rate of viability for long-term culture of yew cells in bioreactor, final cell volume less than 50% in SCV
seemed to be the best. Time course study revealed that 30% of inoculum density was suitable for fed batch culture. Among the
tested conditional media, the rate of 1∶2 (old medium: fresh medium) was recorded at the best for cell growth. 相似文献
30.
Localized reversible frameshift mutation in an adhesin gene confers a phase-variable adherence phenotype in mycoplasma 总被引:13,自引:0,他引:13
The variable adherence-associated (Vaa) antigen of Mycoplasma hominis is an abundant surface lipoprotein adhesin that may mediate important interactions of this wall-less prokaryotic pathogen with the human host. Extensive mutational variation of Vaa size, as well as sequence and antigenic divergence, has been described previously. Using a series of clonal isolates representing an isogenic lineage of variants oscillating in Vaa expression, Vaa is further shown in this study to undergo high-frequency phase variation in expression, which correlated precisely with the ability of M . hominis to adhere to cultured human cells. Although no DNA rearrangements or sequence differences in the 5' regions flanking vaa alleles were detected between Vaa+ and Vaa− variants, intragenic vaa sequences from this lineage revealed an oscillating mutation involving a single nucleotide deletion/insertion in a short tract of adenine residues near the 5' end of the mature Vaa coding sequence, which created a translational frameshift resulting in either a complete Vaa ORF or an in-frame UAG stop codon immediately downstream of the poly-A tract. Evidence for the occurrence of this high-frequency frameshift mutation in vivo was obtained from analysis of PCR-generated vaa sequences amplified from the joint synovial fluid of a patient with M . hominis -associated arthritis, which indicated that Vaa phase variation occurs during M . hominis infection in the natural host. These results identify a distinctive frameshift mutator element in the vaa gene that governs M . hominis adherence and highlight the importance of mutational alteration of primary gene products on the mycoplasma surface as a means of generating and maintaining functional diversity in the host. 相似文献