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101.
Stimulatory GTP-binding Protein (Gs) and adenylate cyclase prepared from bovine brain cortices were co-reconstituted into asolectin vesicles with or without 1000-fold transmembrane Ca2+ gradient. The results showed that both basal activity and Gs-stimulated activity of adenylate cyclase were highest in proteoliposomes with a transmembrane Ca2+ gradient similar to physiological condition (1 M Ca2+ outside and 1 mM Ca2+ inside) and lowest when the transmembrane Ca2+ gradient was in the inverse direction. Such a difference could be diminished following dissipation of the transmembrane Ca2+ gradient by A23187. Comparable conformational changes of Gs in proteoliposomes were also observed when Gs was labeled with the fluorescence probe, acrylodan. These results may indicate that a proper transmembrane Ca2+ gradient is essential not only for higher adenylate cyclase activity but also for its stimulation by Gs. 相似文献
102.
Capsid assembly and involved function analysis of twelve core protein mutants of duck hepatitis B virus. 总被引:6,自引:6,他引:0 下载免费PDF全文
The roles of different regions of the duck hepatitis B virus (DHBV) core protein on viral capsid assembly and related functions were examined. Twelve deletion and insertion mutations which covered 80% of the DHBV C open reading frame were constructed and expressed in Escherichia coli. The N-terminal region (amino acids 3 to 66) of DHBV core protein was important for its tertiary structure and function in E. coli. The expressed core mutants without this region apparently inhibited E. coli growth. The results of transmission electron microscopy of E. coli thin sections, capsid agarose gel, and sucrose gradient sedimentation demonstrated that a few DHBV core mutants with insertion in the N terminus and deletion in the C terminus retained the ability to form core-like particles in E. coli. However, other mutations in most of N-terminal and central regions strongly inhibited the self-assembly ability of DHBV core protein in E. coli. In addition, the mutant with a C-terminal region deletion (amino acids 181 to 228) lost most of the nucleic acid-binding activity of the DHBV core protein. 相似文献
103.
Deletion mapping of the rotavirus metalloprotein NS53 (NSP1): the conserved cysteine-rich region is essential for virus-specific RNA binding. 总被引:9,自引:7,他引:2 下载免费PDF全文
NS53 (NSP1), the gene 5 product of the group A rotaviruses, is a minor nonstructural protein of 486 to 495 amino acids which binds zinc and contains an amino-terminal highly conserved cysteine-rich region that may form one or two zinc fingers. To study the structure-function of the gene 5 product, wild-type and mutant forms of NS53 were produced by using a recombinant baculovirus expression system and a recombinant vaccinia virus/T7 (vTF7-3) expression system. Analysis of the RNA-binding activity of the wild-type NS53 immobilized onto protein A-Sepharose beads with NS53-specific antiserum showed that the protein exhibited specific affinity for all 11 rotavirus mRNAs. The use of short virus-specific RNA probes indicated that NS53 specifically recognizes an element located near the 5' ends of viral mRNAs. Analysis of the RNA-binding activity of deletion mutants of NS53 showed that the RNA-binding domain resides within the first 81 amino acids of the protein and that the highly conserved cysteine-rich region within this region of the protein is essential for the activity. Gel electrophoresis and Western immunoblot analyses of intracellular fractions derived from infected cells revealed that large amounts of NS53 were present in the cytosol and in association with the cytoskeletal matrix. Indirect immunofluorescence analysis of cells programmed to transiently express mutant forms of NS53 using vTF7-3 indicated that the intracellular localization domain resides between amino acids 84 and 176 of NS53. Together, these data show that the RNA-binding domain and the intracellular localization domain lie upstream from the region of NS53 previously determined not to be essential for replication of rotaviruses in cell culture (J. Hua and J. T. Patton, Virology 198:567-576, 1994). 相似文献
104.
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106.
研究了大蒜(Allium sativum L.)中期染色体的超微结构和RNP物质。常规染色表明,大部分染色体内部有低电子密度区,有的染色体中低电子密度区域较大而似孔洞。银染结果也证明了有大小不等的孔洞存在。Bernhard 染色显示,在染色体周边和染色体内部都有RNP分布。用NaOH 处理证明了Bernhard 染色法所显示的深染区确实含有RNA。RNP量的多少与EDTA 的分化时间呈负相关 相似文献
107.
刺果番荔枝中的番荔枝内酯 总被引:4,自引:0,他引:4
从刺果番荔枝(Annona m uricata L.)的种子中分离到3 个单四氢呋喃型番荔枝内酯类化合物,用波谱方法鉴定为海南哥纳香甲素(how iicin A, S13)、乙素(how iicin B, S5)和新化合物4-去氧海南哥纳香乙素(4-desoxyhow iicin B, S2)。 相似文献
108.
109.
马蹄香大小孢子发生及雌雄配子体形成 总被引:1,自引:0,他引:1
马蹄香(Sarumahenryi,Oliv.)花药壁的发育属双子叶型。花粉母细胞减数分裂为同时型,四分体主要为四面体形,少数为左右对称式排列。腺质绒毡层,其细胞可排列为不规则的两层,双核或多核。到单细胞花粉阶段,绒毡层细胞内切向壁上出现许多乌氏体。成熟花粉为2细胞型,圆球状,具单萌发沟。雌蕊6心皮,上部彼此分离、下部联合。倒生胚珠,双珠被,厚球心。胚囊发育蓼型。成熟胚囊为七细胞结构,但两个助细胞退化较早。 相似文献
110.