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31.
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
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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
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The genes that cause a variety of neurologic and neuromuscular disorders have been mapped to the distal region of Xq. In an effort to isolate genes from this area, a regional genomic library of the distal 30% of Xq was constructed from a single metaphase spread by means of laser microdissection and single unique primer-polymerase chain reaction. Using pooled probes of 1000 clones from the genomic library, human brain cDNA libraries were screened for expressed sequences encoded by this region. From the 250,000 cDNA clones screened so far, 10 nonoverlapping sequences that mapped back to the target portion were isolated. The complete nucleotide sequences of these cDNA clones have been determined. Analysis of the sequences indicates that none has significant similarity to previously characterized primate genes. One sequence mapping to Xq27.3-qter contained an open reading frame of 281 amino acids and was expressed in every tissue tested. This gene, as well as others isolated in this manner, may prove to be a candidate gene for heritable disorders mapping to this region. 相似文献
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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
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细胞间粘附分子1的研究进展 总被引:11,自引:0,他引:11
细胞间粘附分子1(ICAM-1),又名CD54,是一种重要的细胞表面粘附分子,属免疫球蛋白超家族.它可与鼻病毒以及整合素家族成员结合,参与炎症,普通感冒,变态反应及移植排斥反应.文章就其细胞分布、表达调节、结构功能、基因工程以及临床应用进行了综述. 相似文献
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用吸收光谱和圆二色谱的方法研究了蜂毒素与嗜血菌紫膜的相互作用机理.通过与三种在结构和电荷上不同的表面活性剂与紫膜的作用相比较,可以年出蜂毒作为带正电荷的分子与同样带正电的表面活性剂DTAB在引起紫膜凝聚方面表现相同;但在对紫膜可见光区的吸收光谱和圆二色谱的影响上却与具有刚性结构的CHAPS相似,表明蜂毒可在紫膜表面以一种刚性较大的构象(如α螺旋)存在,不能进入膜蛋白流水区的深层.另外,从紫膜-Triton-蜂毒混合作用体系的研究中得到如下推测:蜂毒与Triton竞争菌紫质分子周围的结合位点,可排斥位于菌紫质周围的Triton分子.表明蜂毒具有比Triton更强的与菌紫质的亲和力,从而提供了支持蜂毒分子存在与膜蛋白-菌紫质的直接相互作用的有力证据. 相似文献