全文获取类型
收费全文 | 22913篇 |
免费 | 2855篇 |
国内免费 | 5842篇 |
出版年
2024年 | 157篇 |
2023年 | 498篇 |
2022年 | 943篇 |
2021年 | 1281篇 |
2020年 | 1132篇 |
2019年 | 1268篇 |
2018年 | 1110篇 |
2017年 | 932篇 |
2016年 | 1046篇 |
2015年 | 1486篇 |
2014年 | 1864篇 |
2013年 | 1814篇 |
2012年 | 2216篇 |
2011年 | 1975篇 |
2010年 | 1496篇 |
2009年 | 1336篇 |
2008年 | 1536篇 |
2007年 | 1396篇 |
2006年 | 1244篇 |
2005年 | 1074篇 |
2004年 | 933篇 |
2003年 | 870篇 |
2002年 | 791篇 |
2001年 | 587篇 |
2000年 | 496篇 |
1999年 | 422篇 |
1998年 | 253篇 |
1997年 | 237篇 |
1996年 | 176篇 |
1995年 | 170篇 |
1994年 | 125篇 |
1993年 | 100篇 |
1992年 | 84篇 |
1991年 | 76篇 |
1990年 | 69篇 |
1989年 | 77篇 |
1988年 | 44篇 |
1987年 | 38篇 |
1986年 | 38篇 |
1985年 | 20篇 |
1984年 | 23篇 |
1983年 | 27篇 |
1982年 | 38篇 |
1981年 | 20篇 |
1980年 | 8篇 |
1979年 | 10篇 |
1978年 | 6篇 |
1977年 | 8篇 |
1965年 | 6篇 |
1964年 | 6篇 |
排序方式: 共有10000条查询结果,搜索用时 15 毫秒
91.
MethodsforHandy,RapidIsolationofHighQualityRNAbyGuanadiumThioeyauateDuJianYuanYanhuaMaShenglinDongZhiwei(BeijingInstituteforCancerResearchBeijing100034)硫氰胍是一种有效的蛋白变性剂。早在1979年,Chirgwin等就利用氯化铯/硫氰胍超离心技术成功地从RNA酶富集的胰脏组织中提取出未降解的RNA分子[2],从而使它成为抑制RNA酶的首选药物并得到广泛使用,但受到超速离心设备的限制。1983年,Cathala报道了氯化锂/硫氰胍RNA提取法[1]。该方法操作简便,获得的RNA质量很高,但所需时间较长。为了能在短时间内更快… 相似文献
92.
Wen Wang Ai-hua Liu Shi-ying Lin Hong Lan Bing Su De-wen Xie Li-ming Shi 《Biochemical genetics》1994,32(9-10):371-378
mtDNA genotypes of six domestic horses (three adult short horses whose heights are under 1 m and three common domestic horses) from a small region of 15 km2 in Malipo county of Yunnan province of China were investigated by the technique of restriction fragment length polymorphism (RFLP) with 16 restriction endonucleases which recognize 6-bp sequences. An average of 56 fragments for an individual was obtained. Unlike other domestic animals, this population of horses exhibits high mtDNA genetic diversity. Each of the six horses has a specific mtDNA genotype showing a pattern of multiple maternal origins, as suggested by fossil and literature records. We think the population of horses is an amazing seed-resource pool of horses and hence deserves to be paid more attention from the view of conservation genetics. However, it is also remarkable that we did not find any typical mtDNA genetic markers which would discriminate between short horses and common domestic horses. 相似文献
93.
The red alga Porphyra purpurea (Roth) C. Agardh has a life cycle that alternates between shell-boring, filamentous sporophytes and free-living, foliose gametophytes. The significant morphological differences between these two phases suggest that many genes should be developmentally regulated and expressed in a phase-specific manner. In this study, we prepared and screened subtracted complementary DNA (cDNA) libraries specific for the sporophyte and gametophyte of P. purpurea. This involved the construction of cDNA libraries from each phase, followed by the removal of common clones through subtractive hybridization. Sampling of the subtracted libraries indicated that 8–10% of the recombinant colonies in each library were specific for the appropriate phase. Of 20 putative phase-specific cDNAs selected from each subtracted library, eight unique clones were obtained for the sporophyte and seven for the gametophyte. After confirming their phase-specificities by hybridization to gametophyte and sporophyte messenger RNA, these 15 phase-specific cDNAs were sequenced, and the deduced amino acid sequences were used to search protein databanks. Two proteins encoded by the sporophyte-specific cDNAs and two by the gametophyte-specific cDNAs were identified by their similarity to databank entries. 相似文献
94.
Dong-Her Wu Chiou-Yen Wen Long-Liu Lin Wen-Shen Chu Wen-Hwei Hsu 《Letters in applied microbiology》1994,19(2):67-69
The isoamylase activity of Pseudomonas amyloderamosa WU 5315 was stable over the pH range from 5.5 to 6.25 while only about 30% of the activity remained at pH 6.5. Low isoamylase activity (418 U ml-1 ) was produced by the cells grown at high pH. Activity reached almost 3000 U ml-1 when pH was kept below 6.0 during the fermentation. With 1% glucose plus 2% maltose instead of 3% maltose as carbon source, however, no pH control was required and the isoamylase activity of Ps. amyloderamosa WU 5315 increased to 3400 U ml-1 . 相似文献
95.
5e-tert-Butyl-2e-[4-(substituted-ethynyl)phenyl]-1,3-dithianes with selected functional groups (R) on the ethynyl moiety are potent blockers of the GABA-gated chloride channel measured as inhibitor concentration (IC50) for 4-n-[3H]propyl-1-(4-ethynylphenyl)-2, 6,7-trioxabicyclo[2.2.2]octanebinding to bovine brain membranes. The terminal R substituents were introduced by coupling 5e-tert-butyl-2e-(4-iodophenyl)-1,3-dithiane with HC ≡ CR or 5e-tert-butyl-2e-(4-ethynylphenyl)-1,3-dithiane with XR. The potency of the parent compound (R=H) with an IC50 of 21 μM is equaled or exceeded by up to 7-fold (i.e. IC50 = 3–21 μM) by several carboxylic acids [R = (CH2)nCO2H (n = 0–3), (CH2nOCH2CO2H (n = 1–3) and CH2SCH2 CO2H] and their esters and two phosphonic acids (CH2CH2PO3H2 and CH2OCH2PO3H2) but not their esters. These carboxyl and phosphonic acids (and their salts) include the most potent water-soluble chloride channel blockers known. Conversion to the monosulfones increases activity of the R = H and CH2OH analogs by 1.2- to 3-fold but decreases that of the R = CH2CH2CO2R′ (R′ = H or CH3) derivatives by 3- to 13-fold. Quantitative structure-activity analyses for 44 2-[4-(substituted-ethynyl)phenyl]-dithianes suggests that the principal feature of the R substituent for high activity is its polarizable volume modeled as molecular refractivity, i.e. this substituent is not a well-defined pharmacophore and undergoes a structurally non-specific interaction with the receptor. These observations lay the background for preparing candidate affinity probes. 相似文献
96.
以番茄(Lycopersicon esculentum Mill.)雌蕊和幼果的组织作外植体可以诱导果实状结构的再生。这种果实状结构在离体条件下能培养成熟,成熟时具红色。解剖观察表明:果实状结构由果肉和包围在外面的果皮组成,无种子和胎座。外源激素和外植体年龄的试验揭示:1.以雌蕊组织作外植体时,仅附加外源细胞分裂素就可以诱导果实状结构的再生,外源生长素似乎不是必需的,最高的诱导频率(50.0% )出现在仅附加玉米素0.5 m g/L的组合。2.从直径4—12 m m 的幼果上分离的外植体在附加外源激素的培养基上均可诱导果实状结构的再生,但只有从直径8 m m 的果实分离的组织块作外植体并将它们培养在6-BAP 2 m g/L,NAA0.1 m g/L的培养基上时,果实状结构的诱导频率最高(62.5% )。为了探讨在果实状结构再生中表现出来的细胞全能性的表达,提出了植物细胞全能性的部分表达(Partialexpression ofplant celltotipotency)的概念并进行了讨论。 相似文献
97.
双价外壳蛋白基因植物表达载体的构建及马铃薯转基因植株的鉴定 总被引:10,自引:0,他引:10
在克隆了马铃薯X病毒(PVX)、马铃薯Y 病毒(PVY)和马铃薯卷叶病毒(PLRV)的外壳蛋白基因的基础上,构建同时包含PVX和PVY 与PVY 和PLRV 两个外壳蛋白基因植物表达框架的表达载体,通过农杆菌(Agrobacterium tumefaciens)介导转化烟草(Nicotianatabacum )和生产上常用的几个马铃薯(Solanum tuberosum )优良品种:“Favorita”、“虎头”、“克4”。经PCR检测证明外源基因已整合到植物的染色体上,得到批量转基因植株。在转PVX+PVY 外壳蛋白基因的烟草上接种PVX (5 μg/m L)、PVY(20 μg/m L)病毒,得到有一定抗性的植株 相似文献
98.
We have analyzed a total of 12 different global and local multiple
protein-sequence alignment methods. The purpose of this study is to
evaluate each method's ability to correctly identify the ordered series of
motifs found among all members of a given protein family. Four
phylogenetically distributed sets of sequences from the hemoglobin, kinase,
aspartic acid protease, and ribonuclease H protein families were used to
test the methods. The performance of all 12 methods was affected by (1) the
number of sequences in the test sets, (2) the degree of similarity among
the sequences, and (3) the number of indels required to produce a multiple
alignment. Global methods generally performed better than local methods in
the detection of motif patterns.
相似文献
99.
100.
Selection of antibiotic-resistant mutants with enhanced isoamylase activity in Pseudomonas amyloderamosa 总被引:3,自引:0,他引:3
Dong-Her Wu Chiou-Yen Wen Wen-Shen Chu Long-Liu Lin Wen-Hwei Hsu 《Biotechnology letters》1993,15(9):883-888
Summary Isoamylase-hyperproducing strains of Pseudomonas amyloderamosa were bred by mutagenesis with UV light and N-methyl-N-nitro-N-nitrosoguanidine (NTG). The selection criterion for such strains was based on the formation of large turbid zones around the bacterial colonies in agar medium containing antibiotics and 1% waxy corn starch. Mutant WN6410 was obtained by treating P. amyloderamosa JD210 with five cycles of 1 × 104 J UV light and one cycle of NTG. P. amyloderamosa WN6410 had 22-fold increase in isoamylase activity when compared to wild-type strain SB15 and the maximal enzyme activity, 5,100 U/ml, could be achieved within 48 h in 2.5 L fed-batch fermentation. 相似文献