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:从到   被引次数:从到   他引次数:从到 提示：输入*表示无穷大
全文获取类型
  收费全文   17173篇
  免费   1416篇
  国内免费   7篇
专业分类
  18596篇
出版年
  2024年   21篇
  2023年   130篇
  2022年   305篇
  2021年   533篇
  2020年   309篇
  2019年   399篇
  2018年   457篇
  2017年   352篇
  2016年   662篇
  2015年   1073篇
  2014年   1159篇
  2013年   1347篇
  2012年   1630篇
  2011年   1456篇
  2010年   929篇
  2009年   773篇
  2008年   1019篇
  2007年   995篇
  2006年   886篇
  2005年   808篇
  2004年   772篇
  2003年   617篇
  2002年   617篇
  2001年   105篇
  2000年   90篇
  1999年   110篇
  1998年   125篇
  1997年   96篇
  1996年   90篇
  1995年   65篇
  1994年   60篇
  1993年   49篇
  1992年   48篇
  1991年   38篇
  1990年   37篇
  1989年   48篇
  1988年   38篇
  1987年   24篇
  1986年   20篇
  1985年   28篇
  1984年   30篇
  1983年   22篇
  1982年   19篇
  1981年   18篇
  1980年   21篇
  1979年   19篇
  1978年   13篇
  1976年   17篇
  1974年   13篇
  1968年   10篇
排序方式： 共有10000条查询结果，搜索用时 15 毫秒
991.
Pyrrolnitrin is a secondary metabolite derived from tryptophan and has strong antifungal activity. Recently we described four genes, prnABCD, from Pseudomonas fluorescens that encode the biosynthesis of pyrrolnitrin. In the work presented here, we describe the function of each prn gene product. The four genes encode proteins identical in size and serology to proteins present in wild-type Pseudomonas fluorescens, but absent from a mutant from which the entire prn gene region had been deleted. The prnA gene product catalyzes the chlorination of l-tryptophan to form 7-chloro-l-tryptophan. The prnB gene product catalyzes a ring rearrangement and decarboxylation to convert 7-chloro-l-tryptophan to monodechloroaminopyrrolnitrin. The prnC gene product chlorinates monodechloroaminopyrrolnitrin at the 3 position to form aminopyrrolnitrin. The prnD gene product catalyzes the oxidation of the amino group of aminopyrrolnitrin to a nitro group to form pyrrolnitrin. The organization of the prn genes in the operon is identical to the order of the reactions in the biosynthetic pathway.The antibiotic pyrrolnitrin [3-chloro-4-(2′-nitro-3′-chlorophenyl)pyrrole] (PRN) is produced by many pseudomonads and has broad-spectrum antifungal activity (1, 5, 12–14, 17). PRN has been implicated as an important mechanism of biological control of fungal plant pathogens by several Pseudomonas strains (12–14), including P. fluorescens BL915, from which the prn genes were isolated (10).Tryptophan was identified as the precursor for PRN, based on the feeding of cultures with isotopically labeled and substituted tryptophan (2, 7, 8, 17, 25). Biosynthetic pathways were proposed as early as 1967 (7) and have been refined on the basis of tracer studies and the isolation of intermediates (Fig. ​(Fig.1)1) (2, 8, 17, 19, 23, 25). Recently, Hammer et al. (9) described the cloning and characterization of a 5.8-kb DNA region which encodes the PRN biosynthetic pathway. This DNA region confers the ability to produce PRN when expressed heterologously in Escherichia coli and contains four genes, prnABCD, each of which is required for PRN production. In the research described here, we used mutants in which each of the four genes was disrupted and strains which overexpress the individual genes to elucidate the function of each gene product in PRN biosynthesis. Open in a separate windowFIG. 1Biosynthetic pathways for PRN as proposed by van Pée et al. (23) (A) and by Chang et al. (2) (B). The reactions catalyzed by the PRN biosynthetic enzymes encoded by the prnABCD genes are indicated above the appropriate reaction arrows.

Bacterial strains and plasmids.

The bacterial strains and plasmids used in this study are described in Table ​Table1.1. Pseudomonas strains were cultured in Luria-Bertani medium at 28°C. Antibiotics, when used, were added at the following concentrations: tetracycline, 30 μg/ml; and kanamycin, 50 μg/ml. The expression vector pPEH14 consists of the Ptac promoter and rrnB ribosomal terminator from pKK223-3 (Pharmacia, Uppsala, Sweden) cloned into the BglII site of the broad-host-range plasmid pRK290 (4). Ptac is a strong constitutive promoter in Pseudomonas (unpublished data). The PRN biosynthetic genes are the coding regions described by Hammer et al. (9). Each coding region was cloned from the translation initiation codon to the stop codon by PCR with restriction sites added to the ends to facilitate cloning. For prnB, the native GTG initiation codon was changed to ATG. The clones were sequenced after PCR.

TABLE 1

Bacterial strains and plasmids used in this study
P. fluorescens strain or plasmidCharacteristicsSource or reference
Strains
 BL915Wild type10
 BL915ΔORF1Deletion in prnA of BL915, Prn−, Kmr9
 BL915ΔORF2Deletion in prnB of BL915, Prn−, Kmr9
 BL915ΔORF3Deletion in prnC of BL915, Prn−, Kmr9
 BL915ΔORF4Deletion in prnD of BL915, Prn−, Kmr9
 BL915ΔORF1–4Deletion in prnABCD of BL915, Prn−, Kmr9
Plasmids
 pPEH14(prnA)pRK290 carrying Ptac functionally fused to the 1.6-kb prnA coding regionThis study
 pPEH14(prnB)pRK290 carrying Ptac functionally fused to the 1.1-kb prnB coding regionThis study
 pPEH14(prnC)pRK290 carrying Ptac functionally fused to the 1.7-kb prnC coding regionThis study
 pPEH14(prnD)pRK290 carrying Ptac functionally fused to the 1.1-kb prnD coding regionThis study
Open in a separate window

Chemical standards.

7-Cl-d,l-tryptophan (7-CT) was synthesized as described by van Pée et al. (24). Monodechloroaminopyrrolnitrin (MDA) was extracted from cultures of P. aureofaciens and verified as described by van Pée et al. (23). Aminopyrrolnitrin (APRN) was prepared from PRN by reduction with sodium dithionite (22). PRN was synthesized according to the method of Gosteli (6).

Western analysis.

To produce antigen, each prn gene was subcloned into a pET3 vector and transformed into E. coli BL21(De3) (Novagen, Inc., Madison, Wis.). Inclusion bodies were purified from induced cultures with protocols from Novagen. Inclusion body protein (100 μg) was run on a preparative Laemmli polyacrylamide electrophoresis gel, blotted to nitrocellulose filters, and stained with Ponceau S. The major band was excised, solubilized in dimethyl sulfoxide, and used by Duncroft, Inc. (Lovettsville, Va.), to immunize goats and produce antiserum against each PRN protein.Cultures of P. fluorescens BL915 were grown for 48 h in Luria-Bertani medium with the appropriate antibiotics. The cells were pelleted and resuspended in a small volume of Tris-EDTA. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western analysis were performed as described by Sambrook et al. (21). The primary antiserum (goat anti-PRN protein) was diluted 1/1,000, and the secondary antibody (rabbit anti-goat immunoglobulin G conjugated to peroxidase; Pierce, Rockford, Ill.) was diluted 1/3,000. Bands were visualized with an enhanced chemiluminescence kit (Amersham, Arlington Heights, Ill.). This Western analysis demonstrated that each antibody recognized a single protein band from wild-type BL915, and these bands were not present in BL915ΔORF1–4 (Fig. ​(Fig.2).2). The molecular weights of the recognized proteins were consistent with the sizes predicted from the gene sequences. Each prn gene was expressed on a plasmid in BL915ΔORF1–4. In each case, the protein product of the cloned gene reacted only with the expected antibody and was identical in size to the band detected by that antibody in wild-type BL915 (Fig. ​(Fig.2).2). Open in a separate windowFIG. 2Western blot analysis of the protein products of prn genes cloned from P. fluorescens BL915. Individual genes were expressed on plasmids in the host strain BL915ΔORF1–4. BL915 wild-type and BL915ΔORF1–4 controls are included on each blot. Blots A, B, C, and D were probed with antibodies raised against the products of prnA, prnB, prnC, and prnD, respectively. Arrows indicate the positions of the 60- and 42-kDa molecular mass markers.

Intermediate analysis and feeding experiments.

To determine which biosynthetic intermediates were produced by the prn gene deletion mutants, 2-day-old cultures were extracted with an equal volume of ethyl acetate. The organic phase was dried under vacuum, and the residue was dissolved in a small volume of methanol. Thin-layer chromatography (TLC) was performed on silica-coated plates with toluene or hexane-ethyl acetate (2:1) as the mobile phase. PRN, APRN, MDA, and aminophenylpyrrole (APP) were visualized with van Urk’s reagent as described previously (22).To further clarify which biosynthetic step was blocked in each deletion mutant, intermediate feeding experiments were conducted. Cultures (10 ml) were incubated at 28°C for 48 h. Biosynthetic intermediates were dissolved in a small volume of methanol and added to 4 ml of culture at the following final concentrations: 7-CT, 2.5 μg/ml; MDA, 25 μg/ml; APRN, 12.5 μg/ml. The cultures were incubated for an additional 4 h at 28°C and then extracted with ethyl acetate and analyzed by TLC and liquid chromatography-mass spectrometry as described above.MDA, APRN, and PRN were not detected in cultures of BL915ΔORF1 (Fig. ​(Fig.3),3), indicating that this mutant is blocked at an early step in PRN biosynthesis. BL915ΔORF1 was able to produce PRN when 7-CT, MDA, or APRN was supplied exogenously (Table ​(Table2).2). When prnA was expressed in the absence of other prn genes (i.e., in BL915ΔORF1–4), 7-chloro-l-tryptophan (7-CLT) accumulated. The identity of 7-CLT was verified by comparison of results of high-performance liquid chromatography and mass spectra with chemically synthesized 7-CT. These results indicate that the prnA gene product catalyzes the chlorination of l-tryptophan. Open in a separate windowFIG. 3Accumulation of PRN biosynthetic intermediates in P. fluorescens BL915 and prn gene deletion mutants derived from it. Extracts from 2-day-old cultures were separated by TLC on silica plates with hexane-ethyl acetate (2:1 [vol/vol]) as the mobile phase. Metabolites were visualized with van Urk’s reagent. Arrows indicate the positions of MDA (olive green), APRN (reddish brown), and PRN (purple).

TABLE 2

Production of PRN by deletion mutants when supplied with biosynthetic intermediates in the growth medium
StrainResult with intermediate added to culturesa
7-CTMDAAPRN
BL915ΔORF1+++
BL915ΔORF2−++
BL915ΔORF3−−+
BL915ΔORF4−−−
Open in a separate windowa+, PRN detected; −, PRN not detected. Hohaus et al. (11) presented additional evidence of the chlorinating activity of the prnA gene product, specifically, the chlorination of l-tryptophan to form 7-CLT by cell extracts from P. fluorescens strains which expressed the prnA gene, but which did not contain any of the other prn genes. To clarify which isomer was produced, Hohaus et al. (11) extracted 7-CLT from the bacteria and oxidized it to the corresponding indole-3-pyruvic acid with amino acid oxidases. Since the isolated 7-CLT was degraded by l-amino acid oxidase, but not by d-amino acid oxidase (11), it must be in the l configuration. The deduced amino acid sequence for prnA contains a consensus NAD binding site (9), and, indeed, NADH is a required cofactor for the prnA gene product.Cultures of BL915ΔORF2 produced 7-CLT, but 7-chloro-d-tryptophan (11) and other PRN biosynthetic intermediates were not detected (Fig. ​(Fig.3).3). BL915ΔORF2 produced PRN when supplied with exogenous MDA or APRN, but not when supplied with 7-CT (Table ​(Table2).2). When prnB was expressed in strain BL915ΔORF1–4, exogenously supplied 7-CT was converted to MDA (Fig. ​(Fig.4).4). These results indicate that the prnB gene product catalyzes the rearrangement of the indole ring to a phenylpyrrole and the decarboxylation of 7-CLT to convert 7-CLT to MDA. While it is somewhat surprising that a single enzyme carries out both the ring rearrangement and decarboxylation, Chang et al. (2) postulated a mechanism for such a reaction on a single enzyme some 16 years ago. The prnB gene product also catalyzed the production of APP (Fig. ​(Fig.4),4), presumably by using tryptophan as a substrate. Open in a separate windowFIG. 4In vivo conversion of PRN biosynthetic intermediates by the products of single prn genes. Individual genes were expressed on plasmids in the host strain BL915ΔORF1–4, and biosynthetic intermediates were added to the culture medium as indicated. Culture extracts were separated by TLC on silica plates with toluene as the mobile phase. Metabolites were visualized with van Urk’s reagent. Arrows indicate the positions of APP (dark green), MDA (olive green), APRN (reddish brown), and PRN (purple).MDA accumulated in cultures of BL915ΔORF3, but APP, APRN, and PRN were not detected (Fig. ​(Fig.3).3). BL915ΔORF3 was able to produce PRN when supplied with APRN in the culture medium, but not when supplied with 7-CT or MDA (Table ​(Table2).2). Strain BL915ΔORF1–4 expressing prnC converted exogenously supplied MDA to APRN (Fig. ​(Fig.4).4). These data indicate that the prnC gene product catalyzes the chlorination of MDA to form APRN. Cell extracts of the P. fluorescens strain which overexpresses the prnC gene (but does not contain the other prn genes) can also catalyze the chlorination of MDA to form APRN (11).The prnC gene is homologous to the chl gene from Streptomyces aureofaciens, which encodes a chlorinating enzyme for tetracycline biosynthesis (3, 9). Like prnA, the prnC deduced amino acid sequence contains a consensus NAD binding region (9), and NADH is required for the chlorination of MDA (11). While both prnA and prnC encode halogenating enzymes, they show no homology to previously cloned haloperoxidases (9) or to each other. Furthermore, in contrast to haloperoxidases (16), the two NADH-dependent halogenating enzymes in the PRN biosynthesis pathway are substrate specific (i.e., the tryptophan halogenase does not catalyze the chlorination of MDA and vice versa) (11).APRN accumulated in cultures of BL915ΔORF4 (Fig. ​(Fig.3),3), and this mutant was not able to produce PRN when supplied with any of the known PRN biosynthetic intermediates. Strain BL915ΔORF1–4 expressing prnD converted exogenously supplied APRN to PRN (Fig. ​(Fig.4).4). These results indicate that the prnD gene product catalyzes the oxidation of the amino group of APRN to a nitro group forming PRN. In vitro experiments by Kirner and van Pée (15) had suggested that this reaction is catalyzed by a chloroperoxidase; however, gene disruption experiments demonstrated that chloroperoxidases are not involved in PRN biosynthesis in vivo (16). Instead, this oxidation is more likely to be catalyzed by a class IA oxygenase (20), as suggested by the homology of prnD with these enzymes (9).We have shown that each prn gene encodes a protein found in the wild-type BL915 strain and have demonstrated in vivo that these four gene products carry out four biochemical steps which convert l-tryptophan to PRN. None of the conversions were observed in strain BL915ΔORF1–4, from which the entire 5.8-kb prn gene region has been deleted (Fig. ​(Fig.4).4). The arrangement of the genes in the operon is identical to the sequence of reactions in the biosynthetic pathway proposed by van Pée et al. (23) (Fig. ​(Fig.11).Chang et al. (2) proposed an alternate biosynthetic scheme (Fig. ​(Fig.1B)1B) and reported the conversion of exogenously supplied APP to PRN in vivo. Similarly, Zhou et al. (25) reported the conversion of APP to APRN in a cell-free system. These workers concluded that APP is an intermediate in PRN biosynthesis and that ring rearrangement precedes chlorination (Fig. ​(Fig.1B).1B). In the present study, APP accumulated only in strains which overexpressed the prnB gene. Furthermore, APP was not detected in cultures of BL915ΔORF1, which contains functional prnBCD genes expressed from the native promoter, as would be expected if the ring rearrangement (catalyzed by the prnB gene product) occurs before the first chlorination step (catalyzed by the prnA gene product). Like Hamill et al. (8) and van Pée et al. (23), we demonstrated that exogenously supplied 7-CT is converted to PRN. These results, together with the finding that the gene product of prnA catalyzes the NADH-dependent chlorination of l-tryptophan to 7-CLT (11), support the biosynthetic pathway proposed by van Pée et al. (23) (Fig. ​(Fig.1A)1A) and suggest that APP is a side product or dead-end metabolite. Purification and kinetic characterization of the prnA and prnB gene products, including investigations of substrate specificity and regioselectivity, will further clarify the roles of 7-CLT and APP in the PRN biosynthetic pathway.If APP is indeed a dead-end metabolite, it would be advantageous to tightly regulate the amount of prnB gene product present in cells, thus minimizing the diversion of substrate into APP. The prnB gene begins with GTG (9), which is a two- to threefold-less-efficient initiation codon than ATG (18); however, the prnB open reading frame is apparently translationally coupled to the prnA open reading frame (9). Coupling increases translational efficiency and is thought to be a mechanism to ensure coordinate expression of the coupled genes (18). In PRN biosynthesis, translational coupling of prnA and prnB may be a mechanism to regulate the level of prnB gene product present in cells and minimize the diversion of tryptophan to APP.  相似文献   
992.
Mating pair stabilization occurs during conjugative DNA transfer whereby the donor and recipient cells form a tight junction which requires pili as well as TraN and TraG in the donor cell. The role of the outer membrane protein, TraN, during conjugative transfer was examined by introduction of a chloramphenicol resistance cassette into the traN gene on an F plasmid derivative, pOX38, to produce pOX38N1::CAT. pOX38N1::CAT was greatly reduced in its ability to transfer DNA, indicating that TraN plays a greater role in conjugation than previously thought. F and R100-1 traN were capable of complementing pOX38N1::CAT transfer equally well when wild-type recipients were used. F traN, but not R100-1 traN, supported a much lower level of transfer when there was an ompA mutation or lipopolysaccharide (LPS) deficiency in the recipient cell, suggesting receptor specificity. The R100-1 traN gene was sequenced, and the gene product was found to exhibit 82.3% overall similarity with F TraN. The differences were mainly located within a central region of the proteins (amino acids 162 to 333 of F and 162 to 348 of R100-1). Deletion analysis of F traN suggested that this central portion might be responsible for the receptor specificity displayed by TraN. TraN was not responsible for TraT-dependent surface exclusion. Thus, TraN, and not the F pilus, appears to interact with OmpA and LPS moieties during conjugation, resulting in mating pair stabilization, the first step in efficient mobilization of DNA.  相似文献   
993.
The distinct structural properties of heterochromatin accommodate a diverse group of vital chromosome functions, yet we have only rudimentary molecular details of its structure. A powerful tool in the analyses of its structure in Drosophila has been a group of mutations that reverse the repressive effect of heterochromatin on the expression of a gene placed next to it ectopically. Several genes from this group are known to encode proteins enriched in heterochromatin. The best characterized of these is the heterochromatin-associated protein, HP1. HP1 has no known DNA-binding activity, hence its incorporation into heterochromatin is likely to be dependent upon other proteins. To examine HP1 interacting proteins, we isolated three distinct oligomeric species of HP1 from the cytoplasm of early Drosophila embryos and analyzed their compositions. The two larger oligomers share two properties with the fraction of HP1 that is most tightly associated with the chromatin of interphase nuclei: an underphosphorylated HP1 isoform profile and an association with subunits of the origin recognition complex (ORC). We also found that HP1 localization into heterochromatin is disrupted in mutants for the ORC2 subunit. These findings support a role for the ORC-containing oligomers in localizing HP1 into Drosophila heterochromatin that is strikingly similar to the role of ORC in recruiting the Sir1 protein to silencing nucleation sites in Saccharomyces cerevisiae.  相似文献   
994.
Transforming growth factor-β1 (TGFβ1), a major promoter of myofibroblast differentiation, induces α-smooth muscle (sn) actin, modulates the expression of adhesive receptors, and enhances the synthesis of extracellular matrix (ECM) molecules including ED-A fibronectin (FN), an isoform de novo expressed during wound healing and fibrotic changes. We report here that ED-A FN deposition precedes α-SM actin expression by fibroblasts during granulation tissue evolution in vivo and after TGFβ1 stimulation in vitro. Moreover, there is a correlation between in vitro expression of α-SM actin and ED-A FN in different fibroblastic populations. Seeding fibroblasts on ED-A FN does not elicit per se α-SM actin expression; however, incubation of fibroblasts with the anti-ED-A monoclonal antibody IST-9 specifically blocks the TGFβ1-triggered enhancement of α-SM actin and collagen type I, but not that of plasminogen activator inhibitor-1 mRNA. Interestingly, the same inhibiting action is exerted by the soluble recombinant domain ED-A, but neither of these inhibitory agents alter FN matrix assembly. Our findings indicate that ED-A–containing polymerized FN is necessary for the induction of the myofibroblastic phenotype by TGFβ1 and identify a hitherto unknown mechanism of cytokine-determined gene stimulation based on the generation of an ECM-derived permissive outside in signaling, under the control of the cytokine itself.  相似文献   
995.
Epstein-Barr virus (EBV) is a ubiquitous herpesvirus associated with infectious mononucleosis and several tumors. The BARF1 gene is transcribed early after EBV infection from the BamHI A fragment of the EBV genome. Evidence shown here indicates that the BARF1 protein is secreted into the medium of transfected cells and from EBV-carrying B cells induced to allow lytic replication of the virus. Expression cloning identified colony-stimulating factor-1 (CSF-1) as a ligand for BARF1. Computer-assisted analyses indicated that subtle amino acid sequence homology exists between BARF1 and c-fms, the cellular proto-oncogene that is the receptor for CSF-1. Recombinant BARF1 protein was found to be biologically active, and it neutralized the proliferative effects of human CSF-1 in a dose-dependent fashion when assayed in vitro. Since CSF-1 is a pleiotropic cytokine best known for its differentiating effects on macrophages, these data suggest that BARF1 may function to modulate the host immune response to EBV infection.  相似文献   
996.
For over 100 years, a major focus of photobiological studies has been the unicellular flagellate, Euglena gracilis, an organism well suited for such investigations by its special complement of organelles that may be considered an ancient, yet complete “visual” system. The possible photoreceptive roles of the cytoplasmic stigma and the photoreceptor (paraflagellar swelling) of E. gracilis are still under debate, because of conflicting interpretations of the results produced so far by the different research groups working on this microorganism. This article deals with our hypothesis, first put forward in the late 1980s, that rhodopsin-like proteins are responsible for photo-detection and that the paraxial rod is involved in the control of flagellar movements. This hypothesis uses oriented dipole and electroconformational coupling mechanisms as the physical phenomena that produce signal transduction. A model for phototaxis is presented.  相似文献   
997.
Sticking together: Cell adhesion interactions inArabidopsis reproduction   总被引：1，自引：0，他引：1  
We review the role of the extracellular matrix in transducing environmental signals, focusing on adhesion molecules in plants and animals. Plant reproduction is ideal for investigating cell-cell interactions; recently-describedArabidopsis thaliana mutants defective in cell adhesion during reproduction promise to illuminate unique cell signaling mechanisms. The exteneded abstract of a paper presented at the 13th International Symposium in Conjugation with Award of the International Prize for Biology “Frontier of Plant Biology”  相似文献   
998.
The human immunodeficiency virus type 1 (HIV-1) Nef protein is essential for AIDS pathogenesis, but its function remains highly controversial. During stresses such as growth in the presence of copper or at elevated temperature, myristylated Nef is released from yeast cells and, after extended culture in stationary phase, it accumulates in the supernatant as a dense membranous material that can be centrifuged into a discrete layer above the cell pellet. This material is unique to Nef-producing cells and represents a convenient source of Nef that may have application in further biological studies. Within the yeast cell, electron microscopic examination shows that Nef localises in novel, membrane-bound bodies. These data support the evidence for a role of Nef in membrane perturbation and suggest that there may be a similar localisation for myristylated Nef in HIV-1 infected cells.  相似文献   
999.
To examine the role of matrilysin (MAT), an epithelial cell-specific matrix metalloproteinase, in the normal development and function of reproductive tissues, we generated transgenic animals that overexpress MAT in several reproductive organs. Three distinct forms of human MAT (wild-type, active, and inactive) were placed under the control of the murine mammary tumor virus promoter/enhancer. Although wild-type, active, and inactive forms of the human MAT protein could be produced in an in vitro culture system, mutations of the MAT cDNA significantly decreased the efficiency with which the MAT protein was produced in vivo. Therefore, animals carrying the wild-type MAT transgene that expressed high levels of human MAT in vivo were further examined. Mammary glands from female transgenic animals were morphologically normal throughout mammary development, but displayed an increased ability to produce β-casein protein in virgin animals. In addition, beginning at approximately 8 mo of age, the testes of male transgenic animals became disorganized with apparent disintegration of interstitial tissue that normally surrounds the seminiferous tubules. The disruption of testis morphology was concurrent with the onset of infertility. These results suggest that overexpression of the matrix-degrading enzyme MAT alters the integrity of the extracellular matrix and thereby induces cellular differentiation and cellular destruction in a tissue-specific manner.  相似文献   
1000.
Summary Growth rate, morphology, and responsiveness to mitogenic stimuli and pharmacological treatments were evaluated in early and late cell passages derived from the same clone of the widely used MCF-7 human breast adenocarcinoma cell line. Our results indicate dissimilarities between early (E) and late (L) passages for some of the parameters analyzed. The cells that underwent many subcultivations grew faster than the others; both appeared homogeneous in size and shape. The E cells, subcultured for almost 1 yr, displayed higher sensitivity to the mitogenic action of both estradiol, according to the level of estrogen receptor, and insulin-like growth factor-I than did the L cells, kept in culture for more than 10 yr. Cell responsiveness to two drugs, a novel steroid antiestrogen and a polysulfonated distamycin A derivative, was more pronounced in the early cultures only at the longer time of exposure to the higher concentration of the estrogen antagonist. In addition, a drug-induced inhibition of insulin-like growth factor-I binding to its receptor was shown in both E and L cells, the latter being less sensitive than the former when exposed to the antiestrogen. Finally, MCF-7 E and L cells showed similar behavior when drug-induced apoptosis was tested.  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