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DNA from pea and tobacco chloroplasts hybridizes specifically with probes derived from the gene for a membrane component, the a subunit of ATP synthase of the cyanobacterium, Synechococcus 6301. DNA sequence of the hybridizing region of the pea plastid DNA has revealed that it encodes a protein of 247 amino acids relat... |
The RNA1 of tobacco rattle virus (TRV) has been cloned as cDNA and the nucleotide sequence determined of 2 kb from the 3'-terminal region. The sequence contains three long open reading frames. One of these starts 5' of the cDNA and probably corresponds to the carboxy-terminal sequence of a 170-K protein encoded on RNA1... |
We have isolated and sequenced two maize genomic clones that are homologous to the Drosophila hsp70 gene. One of the maize hsp70 clones contains the entire hsp70 coding region and 81 nucleotides of the 5' nontranslated sequence. The predicted amino acid sequence for this maize protein is 68% homologous to the hsp70 of ... |
Maize genomic fragments containing the regulatory and coding regions of a zein gene for a low size class 23-kd protein have been inserted in an interspecific Escherichia coli-Saccharomyces cerevisiae expression vector in different constructions. The presence of the inducible GAL1-10 upstream activation site (UAS) allow... |
Analysis of clones isolated from a cosmid DNA library indicates that the Serratia marcescens chromosome contains at least two genes, chiA and chiB, which encode distinct secreted forms of the enzyme chitinase. These genes have been characterized by inspection of chitinase activity and secreted proteins in Escherichia c... |
Previous experiments have identified a 657-bp restriction fragment in the non-transcribed region of the Drosophila histone gene cluster that is specifically associated with the histone-depleted nuclear scaffold. The remaining fragments of the 5-kb histone repeat were shown to be readily released from the scaffold; henc... |
The DNA sequence of approximately 3.5 kb of the nodulation (nod) region of the Rhizobium leguminosarum symbiotic plasmid pRL1JI was determined. Three open reading frames were identified; genes corresponding to these have been called nodD, nodE and nodF.nodD is adjacent to nodA and is transcribed in the opposite directi... |
Genomic clones of Drosophila melanogaster were isolated from a lambda library by cross-hybridization with the yeast gene coding for the 150-kd subunit of RNA polymerase II. Clones containing a region of approximately 2.0 kb with strong homology to the yeast gene were shown to code for a 3.9-kb poly(A)-RNA. Part of the ... |
We have obtained by Fourier transformed infra-red (FTIR)-spectroscopy BR-K, BR-L and BR-M difference spectra of bacteriorhodopsin regenerated with isotopically labelled retinals. Thereby, we are able to assign reliably the C(14)-C(15) and C=N stretching vibrations of the various intermediates. The lower C(14)-C(15) str... |
In plants, glutamine synthetase (GS) is the enzyme primarily responsible for the assimilation of ammonia into organic nitrogen. In Phaseolus vulgaris a number of isoenzymic forms of GS are found, each of which consists of eight subunits of mol. wt 41 000-45 000. The GS subunits of P. vulgaris have previously been shown... |
Three cDNA clones coding for the contact site A (csA) protein, a cell adhesion molecule of Dictyostelium discoideum, were isolated by screening a cDNA library with monoclonal antibodies. Two of these clones contained the complete coding region for the csA protein of 1542 bp including a sequence of 57 bp coding for the ... |
Acetylcholine is a major excitatory neurotransmitter in the central nervous system of insects. Using DNA probes of the Torpedo nicotinic acetylcholine receptor (AChR) we have isolated two overlapping cDNA clones encoding a putative neuronal AChR protein from the fruitfly, Drosophila melanogaster. The predicted mature p... |
D1 and D2, two chloroplast proteins with apparent mol. wt of 32 000-34 000, play an important role in the photosynthetic reactions mediated by the membrane-bound protein complex of photosystem II (PSII). We have isolated and characterized an uniparental, non-photosynthetic mutant of Chlamydomonas reinhardtii and show t... |
A number of infectious clones of a Californian isolate of the leafhopper-transmitted geminivirus beet curly top virus (BCTV) have been constructed from virus-specific double-stranded DNA isolated from infected Beta vulgaris and used to demonstrate a single component genome. The nucleotide sequence of one infectious clo... |
Using electron microscopic immunocytochemical staining we demonstrate that the product of gene III of cauliflower mosaic virus (CaMV) is associated with viral particles. Furthermore, a fusion protein, expressed in bacteria, consisting of the N-terminus of beta-galactosidase and the complete gene III protein of CaMV sho... |
The genes for the large (rbcL) and small (rbcS) subunits of ribulose-1,5 bisphosphate carboxylase-oxygenase (RuBPCase) from the cyanobacterium Synechococcus PCC 6301, and the rbcS gene of wheat, have been expressed in Escherichia coli in order to study homologous and heterologous enzyme assembly. Synechococcus L subuni... |
An X-ray structure analysis of the photosynthetic reaction centre from the purple bacterium Rhodopseudomonas viridis provides structural details of the pigment-binding sites. The photosynthetic pigments are found in rather hydrophobic environments provided by the subunits L and M. In addition to apolar interactions, th... |
We have cloned and sequenced a single copy gene encoding a ribosomal protein from the ciliate Tetrahymena thermophila. The gene product was identified as ribosomal protein S25 by comparison of the migration in two-dimensional polyacrylamide gels of the protein synthesized by translation in vitro of hybrid-selected mRNA... |
Mitochondria of S-type cytoplasmic male sterile maize contain two linear double-stranded DNA molecules, S1 and S2. Two open reading frames (ORF1 and ORF2) are present in S2 DNA. Fragments from ORF1 were inserted into plasmids to achieve expression in Escherichia coli. Cells transformed with recombinant plasmids produce... |
Antibodies have been raised against a fusion protein containing the 3' region of the coding sequence of the Antennapedia (Antp) gene fused to beta-galactosidase. The distribution of the protein on whole mount embryos and imaginal discs of third instar larvae was examined by immunofluorescence. In young embryos, express... |
The gene suc1 encodes a product which suppresses certain temperature sensitive mutants of the cell cycle control gene cdc2 of Schizosaccharomyces pombe. Mutants in the suc1 gene or over-expression of its product leads to delays in mitotic and meiotic nuclear division. Deletion of the suc1 gene is lethal and generates s... |
The three-dimensional structure of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) from Rhodospirillum rubrum has been determined at 2.9 A resolution by X-ray crystallographic methods. The MIR-electron density map was substantially improved by two-fold non-crystallographic symmetry averaging. The polypeptide ... |
Transcripts of three distinct ribulose-1,5-bisphosphate carboxylase (RuBPC) small subunit (SS) genes account for approximately 90% of the mRNA for this protein in maize leaves. Transcripts of two of them constitute >80% of the SS mRNA in 24-h greening maize leaves. The third gene contribute approximately 10%. Transcrip... |
When vegetative cells of Dictyostelium discoideum were incubated in nutrient medium containing 5 mM caffeine, adenylate cyclase was induced and exceeded control values 30-fold after 6 h of incubation. The effect was dose-dependent and blocked by cycloheximide. Contact site A, a developmentally regulated membrane glycop... |
The effect of nerve growth factor (NGF) on the expression of neurofilament and Thy-1 genes in rat PC12 pheochromocytoma cells was examined at both the transcriptional and post-transcriptional levels. Addition of NGF to cultured PC12 cells produced increases in mRNAs corresponding to the 68 kd neurofilament protein (NF6... |
DNA-protein complexes formed in vitro with isolated DNA-binding proteins from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius were analyzed by electron microscopy. Two of the proteins (10a and 10b) formed specific structures after incubation with DNA. Protein 10a, at low protein concentrations, showed i... |
Most of the dominant alleles of the homeotic gene Antennapedia (Antp) which show a transformation of antennae into legs are associated with large chromosomal inversions. To determine the molecular mechanisms underlying the dominant phenotype, one of the strongest Antp alleles (Antp) was studied in more detail. The muta... |
A cis acting regulatory region has previously been identified 300-500 bp upstream of the Drosophila glue protein gene, Sgs-4. The functional capabilities of this region have now been examined by fusing it to the Drosophila Adh gene and determining the pattern of expression from the fused construct after transformation.... |
The characteristic morphology of many elements of the Drosophila body plan is crucially dependent upon the proper spatial expression of homeotic selector genes. The Deformed locus, which we isolated by virtue of its homology to the homeo box, is a candidate for a homeotic selector in the head region of the developing e... |
A chimaeric gene composed of the 5' upstream region of STLS1, a leaf/stem specifically expressed gene from Solanum tuberosum, and the RNA-coding as well as the 3' downstream region of patatin, the major storage protein of potato tubers, has been transferred into tobacco plants using the Agrobacterium system. The introd... |
Glutamine synthetase is the key enzyme in the assimilation by plants of reduced nitrogen provided from either the soil or fixed symbiotically in association with Rhizobium. We have isolated a number of cDNA clones for soybean glutamine synthetase (GS) from a nodule-cDNA library, using RNA from polysomes immunoprecipita... |
A chimeric gene encoding the alfalfa mosaic virus (AlMV) coat protein was constructed and introduced into tobacco and tomato plants using Ti plasmid-derived plant transformation vectors. The progeny of the self-fertilized transgenic plants were significantly delayed in symptom development and in some cases completely e... |
We have isolated genomic clones encoding 4-coumarate:CoA ligase (4CL), a key enzyme of general phenylpropanoid metabolism, and have analysed the structure and regulation of the genes contained on these clones. Restriction enzyme and sequence analysis indicated that two distinct 4CL genes, Pc4CL-1 and Pc4CL-2, are repre... |
Clones homologous to the 5' region of the Ultrabithorax gene of Drosophila melanogaster have been isolated from D. pseudoobscura, D. funebris and Musca domestica. Regions that encode most of the Ubx protein have been sequenced in all three of these species, and the 5' upstream region has been sequenced in D. funebris t... |
The sulphur-dependent archaebacterium Thermoproteus tenax has a cylindrical cell shape variable in length, but constant in diameter. Its whole surface is covered by a regular protein layer (S-layer). The lattice has p6 symmetry and a lattice constant of 32.8 nm. The three-dimensional reconstruction from a tilt series o... |
The environmentally regulated synthesis of phenylpropanoid natural products was studied by examining the expression of the gene encoding chalcone isomerase (CHI). This enzyme catalyzes a step common to the synthesis of flavonoid pigments and isoflavonoid phytoalexins. A lambdagt11 library was constructed using mRNA fro... |
We describe a phenotypic assay designed to detect excision of the maize controlling element Ac from a selectable marker gene, neomycin phosphotransferase II (NPT II). An NPT II gene which expresses kanamycin resistance in tobacco cells, and contains a unique restriction enzyme site in the untranslated leader region, wa... |
Transcription in lysed barley plastids and Northern slot blot analyses were used to determine the relationship between changes in RNA levels and transcription during plastid development. Transcription in plastids of 4.5-9-day-old dark-grown or illuminated barley seedlings declined up to 10-fold as a function of plant a... |
The reaction centre of the photosynthetic membrane complex photosystem I (PSI) from the thermophilic cyanobacterium Phormidium laminosum was found to crystallize under a range of conditions. The crystallization method, which can occur in the presence of larger detergent molecules than those used previously for the crys... |
The extracellular matrix is involved in the regulation of differentiation and morphogenesis. Here we report the identification of a sea urchin embryonic extracellular matrix protein by means of a monoclonal antibody BL1 (Mab BL1) and the isolation of the protein from basal lamina preparations. In paraffin sections of f... |
The sequence of 4118 nucleotides upstream of the putative initiator codon of the D. melanogaster Ultrabithorax protein was determined. The transcription initiation site for the corresponding mRNA was identified by S1 nuclease mapping and primed extension. It appears that all embryonic RNA products that encode the first... |
Agrobacterium-mediated transfer from a binary vector was used to produce transgenic Nicotiana tabacum plants that expressed coat protein of the plant virus, alfalfa mosaic virus (AMV). Expression levels of the chimeric gene, which was under the control of the cauliflower mosaic virus 19S promoter, were determined in pr... |
An enzyme complex with dihydrofolate reductase (DHFR, E.C.1.5.1.3.) activity was purified to apparent homogenity from wild-carrot cells. The complex has a mol. wt of 286 kd and contains five polypeptide chains of 95, 70, 50, 45 and 26 kd. The DHFR enzyme activity and methotrexate-binding site are on the 45-kd subunit. ... |
In the fruitfly Drosophila melanogaster the recessive mutation lethal (2) giant larvae affects cell growth and cell adhesion properties of neural and imaginal disc cells during embryonic and larval development. Here we report the nucleotide sequence of a cDNA from one of the two major transcripts of the l (2) gl gene c... |
Different wheat germ extracts were tested for the presence of membranes capable of translocating and processing nascent secretory proteins. One lysate was found in which nascent prehuman-placental lactogen (phPL) was translocated and processed to mature human placental lactogen (hPL). Processing was found to occur conc... |
The complete nucleotide sequence of the gene encoding the cytochrome subunit of the photosynthetic reaction centre from the purple bacterium Rhodopseudomonas viridis, and the derived amino acid sequence are presented. The nucleotide sequence of the gene reveals the existence of a typical bacterial signal peptide of 20 ... |
A gene which confers resistance to the herbicide bialaphos (bar) has been characterized. The bar gene was originally cloned from Streptomyces hygroscopicus, an organism which produces the tripeptide bialaphos as a secondary metabolite. Bialaphos contains phosphinothricin, an analogue of glutamate which is an inhibitor ... |
We have investigated the function of the 30 kd protein of tobacco mosaic virus (TMV) by a reverse genetics approach. First, a point mutation of TMV Ls1 (a temperature-sensitive mutant defective in cell-to-cell movement), that causes an amino acid substitution in the 30 kd protein, was introduced into the parent strain,... |
Synthetic oligonucleotide probes were used to clone two loci from the chromosomal DNA of Paracoccus denitrificans that contain the genes for cytochrome c oxidase (cytochrome aa(3)). One locus seems to contain four or five genes probably forming an operon. Two of these code for the oxidase subunits II and III. Three ope... |
Three of the ten acidic ;pathogenesis-related' (PR) proteins known to accumulate in Nicotiana tabacum cv Samsun NN reacting hypersensitively to tobacco mosaic virus, namely -O, -N and -2, have been shown to have 1,3-beta-glucanase (EC 3.2.1.39) activity. By using sera raised against each protein purified to homogeneity... |
The maize 15-Kd zein structural gene was placed under the regulation of French bean beta-phaseolin gene flanking regions. Agrobacterium tumefaciens-mediated transformation was used to insert the chimeric phaseolin-zein gene into the tobacco genome. Transgenic plants synthesized zein in a tissue-specific manner during t... |
We have cloned 215-kb DNA containing the maternal effect region (ME) of the Shaker gene complex (shC) at 16F of the Drosophila X chromosome. Five translocation and deletion breakpoints have been mapped on the cloned DNA allowing a correlation of the genetic map to transcription units. The ME region spans 100 kb. The ge... |
The decision of an ectodermal cell to take on a neural or an epidermal fate depends on its interactions with the neighbouring cells. In Drosophila melanogaster, the available evidence suggests that a regulatory signal necessary for epidermal commitment is built by the products of the so-called neurogenic genes. We have... |
Using Agrobacterium-mediated transformation, two genes for phytohemagglutinin-L (PHA-L), the lectin seed protein of the common bean Phaseolus vulgaris, were stably integrated into the tobacco genome. The two alleles for PHA-L, dlec2 and pdlec2, were obtained from a normal cultivar (Greensleeves) and a lectin-deficient ... |
Two different DNA fragments encoding ornithine carbamoyltransferase (OCTase) were cloned from Pseudomonas syringae pv. phaseolicola NPS3121. These fragments did not cross-hybridize and encoded OCTases which differed with respect to their sensitivity to purified phaseolotoxin, an OCTase inhibitor produced by this phytop... |
The functions of type 1 and 2 carbohydrates of the contact site A (csA) glycoprotein of Dictyostelium discoideum have been investigated using mutants lacking type 2 carbohydrate. In two mutant strains, HG220 and HG701, a 68-kd glycoprotein was synthesized as the final product of csA biosynthesis. This glycoprotein accu... |
It has previously been shown that a region of 100 bp in the Drosophila hsp27 promoter is sufficient to confer ecdysone inducibility on a heterologous gene. We now show, using binding and DNase I footprinting assays, that a 23-bp hyphenated dyad within this sequence forms a protein-binding site, and that this is suffici... |
A wild-type allele of the A1 gene of Zea mays contains a 1.1-kb-long insert termed Cin4-1, which alters the structure of the transcription unit compared to other A1 alleles. The Cin4-1 element is a member of a family of elements occurring in 50-100 copies in the maize genome. Genomic cloning and sequence analysis of se... |
The nuclear-coded 22 kd heat-shock protein (HSP-22) which is transported into the chloroplast and localized in the thylakoids was further characterized and found to be located in the grana lamellae (stacked thylakoids) as an extrinsic protein in the green alga Chlamydomonas reinhardtii. Inhibition of photosynthetic ele... |
The isolation, identification and structure of the spalt gene is described. This novel homeotic gene of Drosophila is required for the establishment of the posterior-most head and the anterior-most tail segments of the embryo. It encodes a small mRNA of 0.8 kb which is under the control of over 15 kb of upstream sequen... |
The translational initiation point for Rhizobium meliloti nifA, the specific activator for nitrogen fixation (nif) genes, was determined. When expressed in an Escherichia coli linked transcriptional-translational system the DNA coding for the R.meliloti nifA gene produced four polypeptide bands of 71 000, 67 000, 62 00... |
The eight major open reading frames (ORFs) of cauliflower mosaic virus (CaMV) have been cloned for in vitro transcription and translation. All the ORFs could be translated. Using antisera against either purified virus or specific gene products, the translation products were screened by immunoprecipitation. The products... |
To study the interaction of the nuclear and chloroplast genomes in the biogenesis of the photosynthetic apparatus, nuclear mutants of Chlamydomonas reinhardtii deficient in photosystem II (PSII) activity were analyzed. Two independently-isolated, allelic nuclear mutants show a pleiotropic reduction in a set of function... |
The 46.8-kd NodC protein of Rhizobium meliloti is a membrane protein, essential for nodule formation. Gene fusions of nodC to a portion of the lambda cI repressor gene were used to define the membrane-anchor domain which is necessary for membrane insertion of the NodC protein into the membrane. The transmembrane orient... |
The photosynthetic water oxidase is composed of 15 polypeptides which are grouped around two functional parts: photosystem II and the catalytic manganese centre. Photochemically driven vectorial electron transfer between the manganese centre and bound plastoquinone causes deprotonation-protonation reactions at opposite... |
We describe the structure and expression of an exceptional bithorax complex mutation, C1. This deletion mutation removes major portions of both the Ultrabithorax (Ubx) and abdominal-A (abd-A) protein coding regions, yet retains many of the functions normally executed by these genes. We show that the ends of the C1 dele... |
When tobacco (Nicotiana tabacum) plants were transferred from the dark to continuous white light, the steady-state mRNA levels transcribed from the nuclear genes encoding chloroplast (GapA and GapB) glyceraldehyde-3-phosphate dehydrogenase increased at least 30- to 50-fold, while the mRNA level for the cytosolic enzyme... |
The geminiviruses are a unique group of higher plant viruses that are composed of twin isometric particles which contain circular, single-stranded DNA. Tomato golden mosaic virus (TGMV), a whitefly-transmitted agent, belongs to the subgroup of geminiviruses whose members possess a bipartite genome. The TGMV A genome co... |
The enzyme acetolactate synthase (ALS) is the target enzyme for the sulfonylurea and imidazolinone herbicides. We describe the isolation and characterization of the ALS genes from two herbicide-resistant mutants, C3 and S4-Hra, of Nicotiana tabacum. There are two distinct ALS genes in tobacco which are 0.7% divergent a... |
Transgenic tobacco plants containing maize gene (Z4) encoding a 23-kd zein protein, which is normally synthesized in the endosperm of maize seeds, were obtained using a modified Ti plasmid vector. Although a polyadenylated transcript homologous to the Z4 gene was present in the seeds of some of these transgenic plants,... |
Transgenic tobacco plants engineered to express either the potato virus X (PVX) coat protein (CP+) or the antisense coat protein transcript (CP-antisense) were protected from infection by PVX, as indicated by reduced lesion numbers on inoculated leaves, delay or absence of systemic symptom development and reduction in ... |
Import of the precursor to 5-enolpyruvylshikimate-3-phosphate synthase (pEPSPS) into chloroplasts is inhibited by the herbicide glyphosate. Inhibition of import is maximal at glyphosate concentrations of >/=10 mum and occurs only when pEPSPS is present as a ternary complex of enzyme-shikimate-3-phosphate-glyphosate. Gl... |
The large subunit of ribulose bisphosphate carboxylase from Anacystis nidulans 6301, and the beta subunit of chloroplast ATP synthase from maize, were fused to the transit peptide of the small subunit of ribulose bisphosphate carboxylase from soybean. These proteins were assayed for post-translational import into isola... |
A novel class of highly abundant polypeptides with antifungal activity has been detected in cell walls of barley leaves. Similar polypeptides known as thionins occur not only in monocotyledonous but also in various dictoyledonous plants. The leaf-specific thionins of barley are encoded by a complex multigene family, wh... |
Despite the prokaryotic origins of chloroplasts, a plant chloroplast tRNA precursor is processed in a homologous in vitro system by a pathway distinct from that observed in Escherichia coli, but identical to that utilized for maturation of nuclear pre-tRNAs. The mature tRNA 5' terminus is generated by the site-specific... |
A viable coat protein deletion mutant of cassava latent virus (CLV) DNA 1 has been isolated, suggesting that this geminivirus might be exploited as a gene replacement vector. An extensive deletion of 727 nucleotides within the coat protein gene renders DNA 1 non-infectious. Chimeric clones have been constructed in whic... |
Maize streak virus (MSV), a geminivirus with a one-component genome, encodes a major coat protein RNA which accumulates in infected plants. Using a maize protoplast cell transient expression system, we have defined and studied the promoter which drives rightward transcription of the RNA encoding the coat protein. We ha... |
We have used the ability of Ac to transpose in tobacco to determine which Ac sequences are required for transposition, using a phenotypic assay for Ac excision from an NPTII gene in which excisions of Ac result in calli resistant to the antibiotic kanamycin (Baker et al., 1987). Here we show that deletion of the Ac DNA... |
The vacuole occupies 25-95% of the plant cell volume and plays an essential role in maintaining cytoplasmic homeostasis of nutrients and ions. Recent patch-clamp studies identified ion channels and electrogenic pumps as pathways for the movement of ions and metabolites across the vacuolar membrane (tonoplast). At high ... |
Although the generation of transgenic plants is now routine, the integration of foreign genetic information has so far been at random sites in the genome. We now present evidence for directed integration into a predicted location in the host plant genome. Protoplasts of transgenic tobacco (Nicotiana tabaccum) plants ca... |
A chimeric oat phytochrome structural gene with an uninterrupted coding region was constructed for expression of the monocot protein in transgenic plants. The structural gene was placed under the transcriptional control of either a light-regulated oat phytochrome promoter or the constitutively active cauliflower mosaic... |
The entire genome of single component geminiviruses such as maize streak virus (MSV) consists of a single-stranded circular DNA of ~2.7 kb. Although this size is sufficient to encode only three average sized proteins, the virus is capable of causing severe disease of many monocots with symptoms of chlorosis and stuntin... |
The transformable cyanobacterium, Synechococcus sp. PCC7942, was used to study the genetics of resistance to the herbicide diuron. In wild-type cells, diuron binds to one of the core proteins, called D1, of photosystem II reaction centres. This binding prevents the transfer of electrons from Q(A), the primary quinone a... |
In French bean (Phaseolus vulgarisL.), the glycine-rich wall protein GRP 1.8 is specifically synthesized in protoxylem tracheary elements of the vascular system. A 494 bp upstream promoter fragment of the gene encoding GRP 1.8 was isolated and translationally fused to the beta-glucuronidase reporter gene. Transgenic to... |
A chimeric gene consisting of 1.3 kb of the 5' regulatory region of a member of the potato proteinase inhibitor II gene family, the coding region of the bacterial beta-glucuronidase (GUS) gene and 260 bp of the proteinase inhibitor II 3'-untranslated region containing the poly(A) addition site was introduced into potat... |
We show that expression of common nodulation genes in Rhizobium meliloti is under positive as well as negative control. A repressor protein was found to be involved in the negative control of nod gene expression. Whereas the activator NodD protein binds to the conserved cis-regulatory element (nod-box) required for coo... |
The 33 kd protein of the photosynthetic oxygen-evolving complex is synthesized in the cytoplasm as a larger precursor and transported into the thylakoid lumen via a stromal intermediate form. In this report we describe a reconstituted system in which the later stages of this import pathway can be studied in isolation. ... |
To obtain the recombinant tumor necrosis factor-alpha converting enzyme (TACE) ectodomain and use it as a selective molecule for the screening of TACE peptide inhibitors, the cDNA coding catalytic domain (T800) and full-length ectodomain (T1300) of TACE were amplified by RT-PCR, and the expression plasmids were constru... |
The mechanisms of increased expression of toll-like receptor 4 (TLR-4) during the formation of foam cells were explored. Low density lipoprotein (LDL) was prepared by density gradient ultracentrifugation and oxidized by incubation with CuCl2. The human monocytic leukemia cell line (THP-1) was cultured in RPMI1640. The ... |
The present study aimed to find out dynamic changes of apoptosis in human cytomegalovirus (HCMV) infected cells and the influence of HCMV infection on activation of caspase-3 and the expression of apoptosis-regulating genes, bcl-2 and fas mRNA. The sequential changes of apoptotic cell rate in high and low MOI (MOI = 2.... |
A mammalian expression plasmid pcDNA3. 1-hCC10 was constructed and identified, then CC10 protein expression in A549 lung cancer cell line was detected. A 273 bp cDNA fragment was amplified from the total RNA of normal lung tissue by using RT-PCR and cloned into expression plasmid cDNA3. 1, and the recombinant plasmid w... |
The HL-60 cells were transfected with chk1 antisense and sense chain, and 24 h later subjected to irradiation. Twenty-four h after irradiation, the changes in the chk1 protein expression was assayed by Western blot, and the cell cycles and apoptosis rate detected by FCM. The irradiated apoptosis sensitivity was increas... |
The antinephritic effect of lipo-prostaglandin E1 prostaglandin E1 incorporated in lipid microspheres was investigated using an experimental model of mesangial proliferative glomerulonephritis (MsPGN). Twenty-two female rats were randomly divided into nephritic group (N, n = 6), lipo-prostaglandin E1 treated group (NL,... |
In order to explore the role of connective tissue growth factor (CTGF) in the pathogenesis of renal tubulointerstitial fibrosis, 48 Wistar rats were randomly divided into sham-operated and unilateral ureteral obstruction (UUO) group. On the postoperative day 1, 3, 7 and 14, the rats were killed and the kidneys were rem... |
The role of protease activated receptor-2 (PAR-2) in the renal tubulointerstitial lesion induced by unilateral ureteral obstruction (UUO) was explored. Mice were sacrificed on the day 1, 3, 5, 7, 10, 14 and 21 after UUO. The expression of PAR-2 mRNA and protein and alpha-smooth muscle actin (alpha-SMA) protein in tubul... |
To investigate the expression of cyclooxygenase-2 (COX-2) in ovarian cancer cell lines, RT-PCR and immunocytochemistry were used to detect the expression of COX-2 in 5 ovarian cancer cell lines. The expression of COX-2 mRNA and protein was detected in all 5 cell lines. It is suggested that COX-2 is expressed in ovarian... |
The anti-tumor effect and mechanism of SEA-Fab' coupled protein on gastric tumor was studied. The target cell Walker-256 was treated with SEA-Fab' synthesized chemically or SEA respectively for 24 h, 36 h or 72 h. PBMC+Walke-256 cells served as controls. The apoptotic index of SEA-Fab' against effector cells was detect... |
The change and the role of MAPK cascade pathway and P53 pathway after liver transplantation were explored. Thirty-four punctured donor liver specimens and 10 normal liver specimens were classified as group A (no rejection, n = 10), group B (mild/moderate acute rejection, n = 10), group C (serious acute rejection, n = 8... |
The effects of cyclosporine A (CsA) on Angiontensin II (Ang II)-induced protein contents, c-fos protein levels and cytosolic Ca2+ level ([Ca2+]i) in cultured cardiomyocytes of neonatal rats were observed. Total protein contents were determined by Bradford method. The expression of c-fos protein was detected by Western ... |
By using RT-PCR and immunohistochemistry, the expressions of transforming growth factor beta2 (TGF-beta2) mRNA, proliferating cell nuclear antigen (PCNA) and fibronection (FN) protein in lens epithelial cells (LECs) of age-related nuclear and cortex cataract were detected and compared. The results of RT-PCR revealed th... |
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