首页 | 本学科首页   官方微博 | 高级检索  
文章检索
  按 检索   检索词:      
出版年份:   被引次数:   他引次数: 提示:输入*表示无穷大
  收费全文   45篇
  完全免费   14篇
  2020年   1篇
  2019年   1篇
  2015年   2篇
  2014年   2篇
  2012年   1篇
  2011年   3篇
  2010年   1篇
  2009年   3篇
  2008年   3篇
  2007年   1篇
  2006年   3篇
  2004年   3篇
  2003年   2篇
  2002年   1篇
  2001年   2篇
  2000年   3篇
  1999年   4篇
  1997年   2篇
  1996年   1篇
  1995年   2篇
  1994年   2篇
  1993年   1篇
  1992年   2篇
  1991年   2篇
  1990年   1篇
  1987年   1篇
  1986年   2篇
  1985年   3篇
  1983年   1篇
  1982年   1篇
  1967年   2篇
排序方式: 共有59条查询结果,搜索用时 33 毫秒
1.
C Yanisch-Perron  J Vieira  J Messing 《Gene》1985,33(1):103-119
Three kinds of improvements have been introduced into the M13-based cloning systems. (1) New Escherichia coli host strains have been constructed for the E. coli bacteriophage M13 and the high-copy-number pUC-plasmid cloning vectors. Mutations introduced into these strains improve cloning of unmodified DNA and of repetitive sequences. A new suppressorless strain facilitates the cloning of selected recombinants. (2) The complete nucleotide sequences of the M13mp and pUC vectors have been compiled from a number of sources, including the sequencing of selected segments. The M13mp18 sequence is revised to include the G-to-T substitution in its gene II at position 6 125 bp (in M13) or 6967 bp in M13mp18. (3) M13 clones suitable for sequencing have been obtained by a new method of generating unidirectional progressive deletions from the polycloning site using exonucleases HI and VII.  相似文献
2.
J Messing  J Vieira 《Gene》1982,19(3):269-276
The strategy of shotgun cloning with M13 is based on obtaining random fragments used for the rapid accumulation of sequence data. A strategy, however, is sometimes needed for obtaining subcloned sequences preferentially out of a mixture of fragments. Shotgun sequencing experiments have shown that not all DNA fragments are obtained with the same frequency and that the redundant information increases during the last third of a sequencing project. In addition, experiments have shown that particular fragments are obtained more frequently in one orientation, allowing the use of only one of the two DNA strands as a template for M13 shotgun sequencing. Two new M13 vectors, M13mp8 and M13mp9, have been constructed that permit the cloning of the same restriction fragment in both possible orientations. Consequently, each of the two strands becomes a (+) strand in a pair of vectors. The fragments to be cloned are cleaved with two restriction enzymes to produce a fragment with two different ends. The insertion of such a fragment into the vector can occur only in one orientation. Since M13mp8 and M13mp9 have their array of cloning sites in an antiparallel order, either orientation for inserting a double-digest fragment can be selected by the choice of the vector.  相似文献
3.
4.
Mucosal surfaces are the entry sites for the vast majority of infectious pathogens and provide the first line of defense against infection. In addition to the epithelial barrier, the innate immune system plays a key role in recognizing and rapidly responding to invading pathogens via innate receptors, such as Toll-like receptors (TLR). Bacterial CpG DNA, a potent activator of innate immunity, is recognized by TLR9. Here, we confirm that local mucosal, but not systemic, delivery of CpG oligodeoxynucleotides (ODN) to the genital tract protects mice from a subsequent lethal vaginal herpes simplex virus type 2 (HSV-2) challenge. Since these effects were so local in action, we examined the genital mucosa. Local delivery of CpG ODN induced rapid proliferation and thickening of the genital epithelium and caused significant recruitment of inflammatory cells to the submucosa. Local CpG ODN treatment also resulted in inhibition of HSV-2 replication but had no effect on HSV-2 entry into the genital mucosa. CpG ODN-induced protection against HSV-2 was not associated with early increases in gamma interferon (IFN-gamma) secretion in the genital tract, and CpG ODN-treated IFN-gamma(-/-) mice were protected from subsequent challenge with a lethal dose of HSV-2. Treatment of human HEK-293 cells transfected with murine TLR9 showed that the antiviral activity of CpG ODN was mediated through TLR9. These studies suggest that local induction of mucosal innate immunity can provide protection against sexually transmitted infections, such as HSV-2 or possibly human immunodeficiency virus, at the mucosal surfaces.  相似文献
5.
6.
A maize (Zea mays L.) endosperm cell culture has been shown to efficiently replicate DNA sequences derived from wheat dwarf virus (WDV), a monopartite monocot geminivirus. To analyze sequences necessary for viral replication and to verify their application for a plant gene expression vector, we have developed a 3.7 kilobase pairs Escherichia coli--plant cell shuttle vector, pWI-11. The p15A origin of replication, functional in E. coli, was introduced into the viral sequences. We have replaced the coding region of the coat protein gene by that of bacterial neomycin phosphotransferase II (NPT II) gene. The resulting NPT II gene fusion can serve as a selectable marker in both plant and E. coli systems. Into a unique cloning site in this pWI-11 vector, we introduced a gene fusion carrying the bacterial beta-glucuronidase (GUS) coding region under control of the cauliflower mosaic virus 35S (CaMV35S) gene promoter and terminator. By transferring these viral sequences into protoplasts derived from maize endosperm cell cultures, we have demonstrated that the plasmid pWI-11 can replicate in maize endosperm cells, that the GUS reporter gene introduced into pWI-11 can be expressed at high level in the transformed cells, and that the replicating viral DNA can be rescued from endosperm cells by transforming E. coli in the presence of kanamycin. The level of GUS gene expression increased progressively in transformed endosperm cells during a prolonged culture period, coinciding with replication of the viral sequences in these cells.  相似文献
7.
8.
The nucleotide sequence of a mitochondrial replicon from maize   总被引:2,自引:0,他引:2  
S R Ludwig  R F Pohlman  J Vieira  A G Smith  J Messing 《Gene》1985,38(1-3):131-138
The 1913-bp maize mitochondrial (mt) plasmid was isolated from a suspension culture of a Black Mexican Sweet maize strain, cloned into M13mp vectors, and sequenced by a unidirectional progressive deletion method. The 1.9-kb extrachromosomal double-stranded circular DNA plasmid was found to contain regions of sequence which in other systems are known to be part of origins of replication (ori). This plasmid could be used as a carrier for chimeric genes and a molecular probe for replication.  相似文献
9.
Gas exchanges and leaf water potential (w) of six-years-old trees of fourteen Prunus amygdalus cultivars, grafted on GF-677, were studied in May, when fruits were in active growing period, and in October, after harvesting. The trees were grown in the field under rain fed conditions. Predawn w showed lower water availability in October compared with May. The lowest w values at midday in May increased gradually afterwards, while in October they decreased progressively until night, suggesting a higher difficulty to compensate the water lost by transpiration. However, relative water content (RWC) measured in the morning was similar in both periods, most likely due to some rainfall that occurred in September and first days of October that could be enough to re-hydrate canopy without significantly increasing soil water availability. The highest net photosynthetic rate (P N) was found in both periods early in the morning (08:00–11:00). Reductions in P N from May to October occurred in most cultivars except in José Dias and Ferrastar. In all cultivars a decrease in stomatal conductance (g s) was observed. Photosynthetic capacity (P max) did not significantly change from spring to autumn in nine cultivars, revealing a high resistance of photosynthetic machinery of this species to environmental stresses, namely high temperature and drought. Osmotic adjustment was observed in some cultivars, which showed reductions of ca. 23 % (Duro d' Estrada, José Dias) and 15 % (Tuono) in leaf osmotic potential (). Such decreases were accompanied by soluble sugars accumulation. The Portuguese cultivar José Dias had a higher photosynthetic performance than the remaining genotypes.  相似文献
10.
Chemical modification studies have been conducted on spinach ferredoxin to determine the nature of the groups on ferredoxin involved in its interaction with its reaction partners. Modification of a limited number (three or four) carboxyl groups or of the single histidine residue resulted in a decreased ability of ferredoxin to participate in NADP photoreduction but not in cytochrome c photoreduction, suggesting that these groups may be involved in interaction with ferredoxin:NADP reductase but are not involved in interaction with the reducing side of Photosystem I. In contrast, modification of amino groups or the single arginine residue on ferredoxin had little effect on the ability of ferredoxin to participate in NADP photoreduction, suggesting these groups are not involved in the interaction of ferredoxin with either ferredoxin:NADP reductase or the reducing side of Photosystem I. Attempts to modify tyrosine residues on ferredoxin resulted in destruction of the iron-sulfur center of the protein.  相似文献
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号