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development of porcine adenovirus-3 as an expression vector.porcine adenovirus-3 (pav-3) was developed as an expression vector using homologous recombination in escherichia coli bj 5183. as a prerequisite, the complete genome of pav-3 was first introduced as a paci restriction fragment into a bacterial plasmid. the plasmid, when paci restricted and transfected into swine testicular cells, produces an infectious virus. the potential of this procedure was demonstrated by the construction of several pav-3 recombinants. part of the e3 region, which is noness ...199910091994
generation of infectious genome of bovine adenovirus type 3 by homologous recombination in bacteria.the widely used technique of generating adenovirus vectors by homologous recombination in mammalian cells is usually not very efficient. this communication describes a simple method of generating a plasmid containing the full-length genome of an adenovirus by homologous recombination in bacteria. following transfection of a suitable mammalian cell line with the full-length adenovirus genome, infectious virus progeny could easily be generated. using this technique the generation of adenovirus rec ...199910092136
lesions and transmission of experimental adenovirus hemorrhagic disease in black-tailed deer fawns.adenovirus infection was the cause of an epizootic of hemorrhagic disease that is believed to have killed thousands of mule deer (odocoileus hemionus) in california during the latter half of 1993. a systemic vasculitis with pulmonary edema and hemorrhagic enteropathy or a localized vasculitis associated with necrotizing stomatitis/pharyngitis/glossitis or osteomyelitis of the jaw were common necropsy findings in animals that died during this epizootic. to study transmission of adenovirus infecti ...199910098637
differential chemokine induction by the mouse adenovirus type-1 in the central nervous system of susceptible and resistant strains of mice.mouse adenovirus-type 1 (mav-1) has recently been shown to cause a fatal hemorrhagic encephalopathy in certain strains of mice whereas other strains are resistant. morbidity is associated with a productive infection of cerebrovascular endothelial cells, resulting in necrosis of the vasculature, infarction, hemorrhage and death within 4 - 6 days. previous studies were not able to define a role for the innate or acquired immune response. in the current study we have addressed the effect of mav-1 o ...199910190691
transcription mapping and characterization of 284r and 121r proteins produced from early region 3 of bovine adenovirus type 3.we established the transcription map of early region (e) 3 of bovine adenovirus 3 (bav-3) by northern blot, s1 nuclease protection assays, cdna sequencing, and rt-pcr analysis. five major classes of mrnas were identified, which shared the 3' ends. four classes of mrnas transcribed from the e3 promoter also shared the 5' end, while one major class of mrna transcribed from the major late promoter contained a tripartite leader sequence at the 5' end. these five transcripts have the potential to enc ...199910191200
adenovirus pneumonia in guinea pigs. 199910197252
[the patterns in the molecular biology of the adenoviruses (the results of a 20-year study at the institute of microbiology and virology of the national academy of sciences of ukraine)].scientific data of the highest importance and priority concerning regularities of structural and functional organization of proteins of adenoviruses capsids and peculiarities of expression of the virus genome are as follows: new antigen determinants of hexon and adenovirus fiber have been discovered, their different nature (conformational or linear) and different orientation, depending on the spatial organization of proteins, have been proved; localization of some epitopes has been determined wi ...199910330872
mucosal immunization of calves with recombinant bovine adenovirus-3: induction of protective immunity to bovine herpesvirus-1.to determine the potential of replication-competent (e3-deleted) bovine adenovirus-3 (bav-3) as a delivery system for vaccine antigens in calves, we evaluated the ability of recombinant bav-3 expressing different forms of of bovine herpesvirus-1 (bhv-1) glycoprotein gd to protect against bhv-1 infection in calves that had pre-existing bav-3 specific antibodies. three- to four-month-old calves, vaccinated intranasally with recombinant bav-3 expressing full-length gd (bav3.e3gd) or a truncated ver ...199910355773
biodegradable alginate microspheres as a delivery system for naked dna.sodium alginate is a naturally occurring polysaccharide that can easily be polymerized into a solid matrix to form microspheres. these biodegradable microspheres were used to encapsulate plasmid dna containing the bacterial beta-galactosidase (lacz) gene under the control of either the cytomegalovirus (cmv) immediate-early promoter or the rous sarcoma virus (rsv) early promoter. mice inoculated orally with microspheres containing plasmid dna expressed lacz in the intestine, spleen and liver. ino ...199910369574
experimental inoculation of heifers with bovine adenovirus type 3.nine 2-year-old heifers having bad3-neutralizing antibody titers between 1:120 and 1:1080 were individually exposed intranasally to an aerosol of 10(8) pfu of wild type (wt) bovine adenovirus type 3 (bad3). four animals were kept as non-inoculated controls. the heifers were examined daily for rectal temperature, weight gain/loss, nasal and ocular discharges, and other clinical signs for 10 d post-inoculation. none of the animals showed any sign of clinical disease. virus excretion was observed i ...199910369575
ovine adenovirus vectors overcome preexisting humoral immunity against human adenoviruses in vivo.recombinant human adenoviruses (had) have become widely used as tools to achieve efficient gene transfer. however, successful application of had-derived vectors in clinical trials is limited due to immunological and potential safety problems inherent in their human origin. in this study, we describe a recombinant ovine adenovirus (oav) as an alternative vector for gene transfer in vivo. in contrast to an had vector, the oav vector was not neutralized by human sera. an oav vector which contained ...199910400791
transcription units of e1a, e1b and pix regions of bovine adenovirus type 3.the major mrna species in the e1 region of the genome of bovine adenovirus type 3 (bav3) have been defined by using a combination of pcr, 5' race, northern analysis and dna sequencing. independent transcription initiation sites were identified for each of the e1a, e1b and protein ix (pix) transcription units, but all mrna species terminated at the same poly(a) addition site immediately downstream of the pix open reading frame. thus, the bav3 e1 region, which consists of the e1a and e1b genes tog ...199910423142
transcription map and expression of bovine herpesvirus-1 glycoprotein d in early region 4 of bovine adenovirus-3.early region 4 (e4) of bovine adenovirus type 3 (bav-3) was analyzed by northern blotting, rt-pcr analysis, cdna sequencing, and s1 nuclease protection assays. the transcriptional map of the e4 region of bav-3 has marked dissimilarities from those of mouse adenovirus-1, ovine adenovirus-287, and human adenovirus-2, for which the transcriptional maps have been constructed. the e4 region of bav-3, located between 98.6 and 89.8 mu transcribes seven distinct classes of bovine adenovirus type 3 mrna. ...199910441562
genomic location and nucleotide sequence of a serotype 3 porcine adenovirus hexon gene.the putative hexon gene of a porcine adenovirus serotype 3 (pav3) has been identified, cloned and the nucleotide sequence determined. the genomic location of the pav3 hexon gene was determined and an open reading frame (orf) encoding a polypeptide of 939 amino acids identified. comparison of the nucleotide sequence of the putative pav3 hexon gene with the sequence of the hav2 hexon gene returned an overall identity of approximately 63%. a stop codon 144 nucleotides upstream and a start codon 18 ...199910446655
the adenovirus that causes hemorrhagic disease of black-tailed deer is closely related to bovine adenovirus-3.dna sequence data was obtained from an adenovirus previously shown to be the cause of a distinctive, fatal hemorrhagic disease of black-tailed deer in california. a 256 base fragment of the viral hexon gene was amplified by pcr from purified adenovirus preparations. the amplicon then was cloned and sequenced. phylogenetic relationships with other mammalian adenoviruses were also determined. although sequence analysis of this portion of the hexon gene indicates that the black-tailed deer adenovir ...199910470262
replication-defective bovine adenovirus type 3 as an expression vector.although recombinant human adenovirus (hav)-based vectors offer several advantages for somatic gene therapy and vaccination over other viral vectors, it would be desirable to develop alternative vectors with prolonged expression and decreased toxicity. toward this objective, a replication-defective bovine adenovirus type 3 (bav-3) was developed as an expression vector. bovine cell lines designated vido r2 (hav-5 e1a/b-transformed fetal bovine retina cell [fbrc] line) and 6.93.9 (madin-darby bovi ...199910516020
the role of the l4 33k gene in adenovirus infection.the late phase of adenovirus infection is characterized not only by the synthesis of late proteins and the assembly of new virions, but also by the inhibition of early gene expression and host cell translation. previous work has demonstrated that both of these inhibitory effects depend upon expression from the major late transcription unit (mltu), controlled by the major late promoter (mlp). furthermore, the repression of early gene expression has been shown to be mediated in trans, suggesting a ...199910544122
photodynamic therapy: tumor targeting with adenoviral proteins.a brief summary of the mechanisms involved in photodynamic therapy (pdt) and the role of delivery vehicles for photosensitizer targeting is addressed. phthalocyanines (pc) have been coupled to adenovirus type 2 capsid proteins including the hexon, the penton base and the fiber to enhance their target selectivity. adenovirus penton base proteins contain the arginine-glycine-aspartic acid peptidic sequence (rgd) motif known to bind with great affinity and high specificity to integrin receptors, ex ...199910546549
csiro's 'natural' vaccines. 199910561787
novel human gene transfer vectors: evaluation of wild-type and recombinant animal adenoviruses in human-derived cells.major disadvantages of human adenovirus (had) vectors in gene therapy include preexisting or induced immune responses, and possible coreplication of recombinant had in the presence of wild-type hads. these disadvantages may be overcome by using nonhuman, animal adenoviruses (aads). we evaluated four different aads for their potential use as viral vectors. the canine adenovirus type 2 (cav2) and bovine adenovirus type 3 (bav3) appeared to be suitable systems, as they infect human cells. cav2, but ...199910566887
porcine adenovirus-3 as a helper-dependent expression vector.porcine adenovirus has been proposed as a potential vector for generating novel and effective vaccines for pigs. as a prerequisite for the generation of helper-dependent porcine adenovirus-3 (pav-3) vectors, two e1-complementing porcine cell lines expressing e1 proteins of human adenovirus-5 (hav-5) were made. these cell lines could be efficiently transfected with dna and allowed the rescue and propagation of a pav-3 recombinant, pav201, containing a 0.597 kb e3 deletion and a 0.803 kb e1a delet ...199910580052
the distribution and kinetics of polyomavirus in lungs of intranasally infected newborn mice.the primary cell types that sustain polyomavirus (py) replication following intranasal infection as well as the nature of the host cellular response to py were unknown. as this is an essential and specific site for virus entry, it seems likely that viral gene function must be adapted to these mucosal tissues. using immunohistochemistry and in situ hybridization, we determined the cell types in the lung that support py gene expression and replication following intranasal inoculation of newborn mi ...200010612660
adenovirus pneumonia in guineapigs: an experimental reproduction of the disease.the recently described virus-induced pneumonia in guineapigs (naumann et al., 1981) was experimentally reproducible in newborn animals, though not in preadult animals. baby hamsters and newborn rats were also not susceptible to infection. 10 of 11 infected newborn guineapigs developed pathological changes identical with those found in spontaneous cases. the incubation period was from 5 to 10 days. the agent could not be cultivated in vitro, and therefore no applicable serological tests could be ...198410628789
normal development, wound healing, and adenovirus susceptibility in beta5-deficient mice.integrins have been shown to play important roles in embryonic development, wound healing, metastasis, and other biological processes. alphavbeta5 is a receptor for rgd-containing extracellular matrix proteins that has been suggested to be important in cutaneous wound healing and adenovirus infection. to examine the in vivo function of this receptor, we have generated mice lacking beta5 expression, using homologous recombination in embryonic stem cells. mice homozygous for a null mutation of the ...200010629031
dna sequencing and phylogenetic analysis of the protease gene of ovine adenovirus 3 suggest that adenoviruses of sheep belong to two different genera.until now, the only published ovine adenovirus dna sequence was the complete genome of ovine adenovirus isolate 287 (oav287) which, compared to other mammalian adenoviruses, possesses strikingly unique genomic organisation and should properly be classified into a new adenovirus genus. the protease gene sequence of ovine adenovirus type 3 (oadv-3) was determined and analysed. the results of phylogenetic analysis of the 205 residue long protein demonstrated that oadv-3 belongs to the genus mastade ...200010653919
an ovine adenovirus vector lacks transforming ability in cells that are transformed by ad5 e1a/b sequences.adenoviruses of the mastadenovirus and aviadenovirus genera are able to transform certain cell types and induce tumor formation in susceptible animals. for the mastadenoviruses the e1a/b sequences are largely responsible for these properties but e4 sequences may also be involved. the transforming sequences of the aviadenoviruses, which lack e1a/b and e4 homologues, have not yet been fully identified. the recent proposal for a third genus of adenoviruses, which apparently lack an e1a homologue an ...200010772988
sequence analysis of porcine adenovirus type 3 e1 region, pix and piva2 genes, and two novel open reading frames.the porcine adenovirus type 3 (pad3) genome between map units 0 and 13.7 was sequenced and compared with similar regions of other adenoviruses. this region consists of the left inverted terminal repeat sequences involved in dna packaging, the entire early region 1 (e1) and the protein ix (pix) transcription unit. the lower strand contains the c-terminal end of iva2 of the e2a transcriptional unit and two novel open reading frames (orfs). the e1 transcription unit consists of orfs for proteins ho ...200010773731
serologic detection of adenoviral hemorrhagic disease in black-tailed deer in california.an enzyme-linked immunosorbent assay (elisa) and a serum neutralization (sn) test were developed to measure serum antibodies against the adenovirus causing hemorrhagic disease in free-ranging and captive experimentally-infected black-tailed deer (odocoilenus hemionus columbianus) in california (usa). there was a strong (rho = 0.874) and significant (p < 0.0001) correlation between elisa and sn titers, although the sn assay was more sensitive than the elisa.200010813622
circumvention of vector-specific neutralizing antibody response by alternating use of human and non-human adenoviruses: implications in gene therapy.to determine whether non-human adenovirus-specific antibodies are cross-neutralizing, rabbit and mouse anti-human adenovirus type 5 (had5), anti-bovine adenovirus type 3 (bad3), and anti-porcine adenovirus type 3 (pad3) sera were used in cross-virus neutralization assays. adenovirus neutralizing antibodies were found to be virus-specific, suggesting that virus neutralizing epitope differs significantly in had5, bad3, and pad3. to further investigate whether immunity to an had5-derived vector cou ...200010873758
serological evidence of adenovirus infection in cats.the widespread presence of adenoviruses in various species makes it probable that infection and the carrier state also exist in cats. on the basis of these considerations, investigations were carried out to find antibodies against adenovirus in sera from different cat populations kept under different conditions. for the antibody detection, purified adenovirus was used in an indirect elisa. to produce positive serum, spf cats were immunized with a purified hexon preparation. altogether 632 field ...200010881688
mouse adenovirus type 1 replication in vitro is resistant to interferon.the effects of mouse interferon (ifn)-alpha/beta and recombinant ifn-gamma on mouse adenovirus type 1 (mav-1) replication were investigated in single-cycle infectious virus yield reduction assays on mouse l929 cells. viral yields at 3 days postinfection indicated that wt mav-1 and pme314, an early region 3 null mutant, were relatively insensitive to both ifn-alpha/beta and ifn-gamma, whereas early region 1a (e1a) mutants pme109 (null), dle105 (conserved region 1 deletion, cr1 delta), dle102 (cr2 ...200010936102
sequence analysis and deletion of porcine adenovirus serotype 5 e3 region.a 3000 basepair (bp) region corresponding to the e3 region, the flanking pviii and part of the fiber protein genes, of the two prototype strains (hnf-61 and hnf-70) of porcine adenovirus serotype five (padv-5) was sequenced. a potential e3 promoter and poly-a signals were identified. the size of the e3 region was 2039 (strain hnf-61) and 2020 bp (strain hnf-70) the largest e3 so far reported among padvs. three open reading frames (orf2-4) were identified within the e3 region. based on the predic ...200010958982
dna sequence of frog adenovirus.the genome of frog adenovirus (fradv-1) was sequenced and found to be the smallest of all known adenovirus genomes. the sequence obtained was 26163 bp in size and contains a substantial direct repeat near the right terminus, implying that it was derived by recombination from a parental genome of only 25517 bp. the closest relative of fradv-1 proved to be turkey adenovirus 3, an avian adenovirus with no previously known near relative. sequence comparisons showed that the two viruses have equivale ...200010993931
evaluation of the efficacy of 2',3'-dideoxycytidine against adenovirus infection in a mouse pneumonia model.the antiviral activity of 2',3'-dideoxycytidine (ddc) has been investigated in a mouse pneumonia model. consolidation of lung, histopathological changes, dna synthesis as well as levels of tnfalpha were assayed. in this in vivo model, the oral administration of ddc twice daily over 4 days, displayed an inhibitory effect. the drug significantly reduced histopathologic responses. analysis indicated that under treatment pulmonary lesions were less severe than those of untreated controls. these data ...200010996395
ovine adenovirus vectors mediate efficient gene transfer to skeletal muscle.ovine adenovirus (oav) vectors represent a promising tool for human gene therapy since these vectors overcome the problem of pre-existing immunity against human adenovirus vectors. in this report we investigated the in vivo characteristics of this novel vector system with respect to its potential for gene transfer into skeletal muscle. we found that moderate doses of an oav-derived vector expressing the human alpha1-antitrypsin gene (oavhaat) infected skeletal muscle in mice very efficiently res ...200011001369
adenoviral vector-mediated rescue of the omp-null phenotype in vivo.the use of gene deletion by homologous recombination to determine gene or protein function has wide application in vertebrate neurobiology. an ideal complement to gene deletion would be subsequent gene replacement to demonstrate re-acquisition of function. here we used an adenoviral vector to replace the olfactory marker protein (omp) gene in olfactory receptor neurons of adult omp-null mice and demonstrated the subsequent re-acquisition of function. our results show that short-term expression o ...200011036268
blood clearance rates of adenovirus type 5 in mice.persistence of adenovirus type 5 in blood has implications for the pathogenicity of the virus infection and for the use of this virus in oncolysis and gene therapy. in this study, the kinetics of adenovirus clearance from blood in mice has been evaluated. after a single inoculation of concentrated virus into the vena cava, virus half-life was less than 2 min. depletion of kupffer cells (kc) resulted in increased viraemia. after tail-vein injection, virus and latex beads co-localized within kc. a ...200011038370
adenoviral gene transfer of nitric oxide synthase increases cerebral blood flow in rats.depletion of nitric oxide may play a role in the development of vasospasm after aneurysmal subarachnoid hemorrhage. replenishment of nitric oxide might be a useful treatment for vasospasm. using rats, we performed intracisternal injections of replication-defective adenovirus containing the endothelial nitric oxide synthase (enos) gene and determined the localization of and effect on cerebral blood flow of transgene expression.200011063115
identification and sequence analysis of the core protein genes of bovine adenovirus 2.the dna sequence of the genome of bovine adenovirus type 2 (badv-2) was determined between map units 42.5 and 50. by sequence analysis and homology search, the genes of five structural proteins were identified within this region: the penton base protein (iii; partial sequence), the major core protein precursor (pvii), the minor core protein (v), the mu core protein precursor (px) and the hexon associated protein precursor (pvi; partial sequence). the putative polypeptides were compared to their ...200011074122
optimization of bovine coronavirus hemagglutinin-estrase glycoprotein expression in e3 deleted bovine adenovirus-3.adenoviral vectors expressing foreign genes have many desirable properties in applications such as vaccination. recently, we have generated replication-competent (e3 deleted) bovine adenovirus-3 (bav-3) recombinants expressing significant amounts of glycoprotein d (gd) of bovine herpesvirus-1 (a dna virus). however, attempts to express the rna virus genes using the same strategy were not successful. in an effort to optimize the expression, we have constructed several bav-3 recombinants carrying ...200011074126
effect of adenovirus type 1 and influenza a virus on streptococcus pneumoniae nasopharyngeal colonization and otitis media in the chinchilla.considerable evidence has implicated respiratory tract virus potentiation of bacterial adherence, colonization, and superinfection as a significant factor contributing to the pathogenesis of otitis media (om). influenza a and b viruses, adenovirus, and respiratory syncytial virus are the primary respiratory tract viruses associated with this disease. investigations have established a dramatic increase in the development of experimental om in chinchillas co-inoculated with influenza a virus and s ...200011089992
construction and characterization of recombinant porcine adenovirus serotype 5 expressing the transmissible gastroenteritis virus spike gene.five recombinant porcine adenoviruses of serotype 5 (padv-5) carrying the full-length or the 5' 2.2 kb half of the transmissible gastroenteritis virus (tgev) spike (s) gene were generated by homologous recombination in e. coli strain bj5183 cells and subsequent transfection of swine testicle cells. the foreign genes were inserted into the e3 region of padv-5. one recombinant virus had no deletion in the e3 region, whereas a 1.2 kb fragment was removed from the e3 region in the remainder of the r ...200111125171
the complete nucleotide sequence of porcine adenovirus serotype 5.the complete nucleotide sequence of porcine adenovirus serotype 5 (padv-5) has been determined and the putative genomic map was constructed. the size of the genome was found to be 32621 nucleotides. twenty-eight putative orfs were identified by their homology to other adenovirus or other virus and eukaryotic genes. several special protein sequence motifs were identified by their homology to similar protein motifs. the putative promoter regions, polyadenylation and splice sites were predicted and ...200111172093
alternate adenovirus type-pairs for a possible circumvention of host immune response to recombinant adenovirus vectors.with the help of monoclonal antibodies the existence of at least 18 different earlier not known intertype (it) specific epitopes were demonstrated in different numbers and combinations on the hexons of different adenovirus serotypes. the it specific epitopes play an important role in the experimental gene therapy and in the recombinant adenovirus vaccination because of the harmful immune response of the recipient organisms directed against the many different epitopes of the adenovirus vector. fo ...200111233691
four new inverted terminal repeat sequences from bovine adenoviruses reveal striking differences in the length and content of the itrs.the inverted terminal repeat (itr) of the genome of four bovine adenovirus (badv) types have been sequenced, analysed and compared to the itrs of other adenoviruses. the length of itrs of the examined badvs ranged between 59 and 368 base pairs, thus the presently known longest adenovirus itr sequence is from badv-10. the conserved motifs and characteristic sequence elements of the itrs providing different binding sites for replicative proteins of viral and cellular origin seemed to be distribute ...200111324754
determination of bovine adenovirus-3 titer based on immunohistochemical detection of dna binding protein in infected cells.dna sequence coding for a portion of dna binding protein (amino acids 3-58) of bovine adenovirus type-3 (bav-3) was cloned and expressed in escherichia coli as a fusion protein with schistosoma japonicum glutathione s-transferase. the fusion protein was affinity purified and used to immunize rabbits. immunoprecipitation and western blot analysis showed that the antiserum could specifically recognize a protein of 48 kda in bav-3-infected cells, which was produced both in early and late phases of ...200111337049
characterisation of the fiber gene and partial sequence of the early region 4 of bovine adenovirus 2 (short communication).the full sequence of the fiber gene and partial sequence of the putative 17 kd protein gene of bovine adenovirus-2 (badv-2) were determined. the size of the fiber gene of badv-2 proved to be 561 amino acids, of which the amino acids 37 to 385 form a typical shaft domain of 22 repetitive motifs. on the complementary strand, a gene homologous to the 17 kd protein coded in the e4 region of several human adenoviruses was found. the sequence analysis seems to confirm the presence of an intron in the ...200111402654
comparison of the genome of ovine adenovirus types 1 through 5 by restriction enzyme analysis and dna hybridisation.the dna of the prototype strains of ovine adenovirus (oadv) 1 through 5 was analysed by restriction enzyme (re) digestion. the re patterns generated by hindiii and psti enzymes were characteristic of the examined strains. oadv-2 and 3 resembled each other the most, and their ecori and hindiii patterns seemed to be identical. considering the number of comigrating fragments, serotypes oadv-2, 3, 4 and 5 looked more closely related to each other than to oadv-1. this finding was strengthened by sout ...200011402664
porcine adenoviruses: an update on genome analysis and vector development.although porcine adenoviruses (padv) are present in the swine populations worldwide, they usually do not cause any disease, or the infection is only manifested in a mild diarrhoea or respiratory signs. the importance of adenoviruses, however, is constantly growing as there is a possibility of developing them into viral vector vaccines against more significant swine pathogens. a short summary of the well-established facts of porcine adenoviruses is given and recent developments of the genetic ana ...200011402666
gene transfer by viral vectors into blood vessels in a rat model of retinopathy of prematurity.to test the feasibility of gene transfer into hyaloid blood vessels and into preretinal neovascularisation in a rat model of retinopathy of prematurity (rop), using different viral vectors.200111466260
characterization of a new adenovirus isolated from black-tailed deer in california.an adenovirus associated with systemic and localized vascular damage was demonstrated by transmission electron microscopy and immunohistochemistry in a newly recognized epizootic hemorrhagic disease in california black-tailed deer. in this study, we describe the cultural, physicochemical and serological characteristics of a virus isolated from lung using neonatal white-tail deer lung and turbinate cell cultures. the virus had the cultural, morphological and physicochemical characteristics of mem ...200111504424
bovine adenovirus type 3 e1b(small) protein is essential for growth in bovine fibroblast cells.in order to study the function of bovine adenovirus type 3 (bav-3) e1a and e1b(small) proteins, we constructed two mutants: (a) bav102a carries an in-frame deletion in the coding region for the e1a protein (nt 831-1080); (b) bav102b carries an insertion of triple stop codons in the e1b region (nt 1654, 178 bp downstream of the e1b(small) start codon), which stops the translation of the e1b(small) gene. bav102a virus could grow to the wild-type bav-3 titer in transformed cell line vido r2 (hav-5 ...200111601898
characterization of a cdna encoding the bovine coxsackie and adenovirus receptor.non-human adenoviruses such as bovine adenovirus type 3 (bav-3) that do not replicate in human cells but can infect human cells in culture could provide an attractive alternative to human adenoviral vectors for gene therapy. in addition, a large-animal model for genetic diseases can be very useful for the assessment of the efficacy of adenovector-mediated gene delivery in man. recombinant human subgroup c adenovectors use the coxsackie and adenovirus receptor (car) to enter their target cells. t ...200111688979
insulin gene transcription is mediated by interactions between the p300 coactivator and pdx-1, beta2, and e47.pancreatic beta-cell-type-specific expression of the insulin gene requires both ubiquitous and cell-enriched activators, which are organized within the enhancer region into a network of protein-protein and protein-dna interactions to promote transcriptional synergy. protein-protein-mediated communication between dna-bound activators and the rna polymerase ii transcriptional machinery is inhibited by the adenovirus e1a protein as a result of e1a's binding to the p300 coactivator. e1a disrupts sig ...200211756538
molecular confirmation of an adenovirus in brushtail possums (trichosurus vulpecula).partial genome characterisation of a non-cultivable marsupial adenovirus is described. adenovirus-like particles were found by electron microscopy (em) in the intestinal contents of brushtail possums (trichosurus vulpecula) in new zealand. using degenerate pcr primers complementary to the most conserved genome regions of adenoviruses, the complete nucleotide sequence of the penton base gene, and partial nucleotide sequences of the dna polymerase, hexon, and pvii genes were obtained. phylogenetic ...200211864751
mutational analysis of early region 4 of bovine adenovirus type 3.the primary objective of characterizing bovine adenovirus type 3 (bav3) in greater detail is to develop it as a vector for gene therapy and vaccination of humans and animals. a series of bav3 early region 4 (e4) deletion-mutant viruses, containing deletions in individual e4 open reading frames (orf) or combinations of orfs, were generated by transfecting primary fetal bovine retinal cells with e4-modified genomic dna. each of these mutants was further analyzed for growth kinetics, viral dna accu ...200111883000
cancer gene therapy: fringe or cutting edge?direct targeting of cancer cells with gene therapy has the potential to treat cancer on the basis of its molecular characteristics. but although laboratory results have been extremely encouraging, many practical obstacles need to be overcome before gene therapy can fulfil its goals in the clinic. these issues are not trivial, but seem less formidable than the challenge of killing cancers selectively and rationally--a challenge that has been successfully addressed.200111905804
comparison of the deduced amino acid sequence of guinea pig adenovirus hexon protein with that of other mastadenoviruses.we sought to isolate, clone, and determine the nucleic acid sequence of the guinea pig adenovirus (gpadv) hexon gene. from this, the amino acid sequence of the cloned portion was deduced and compared with a set of mastadenovirus hexons.200111922174
preparation of ovine adenovirus vectors. 200211987792
sequence analysis of porcine adenovirus serotype 5 fibre gene: evidence for recombination.the nucleic acid and deduced amino acid sequence of the fibre gene of the hnf-61 strain of porcine adenovirus serotype 5 (padv-5) was determined and compared to that of the hnf-70 strain of the same serotype (nagy et al., j gen virol 82, 525-529, 2001) and also to adenovirus fibre genes from the genera mastadenovirus and atadenovirus. the putative hnf-61 and hnf-70 proteins were similar to each other, with 90% amino acid identity. conserved amino acid sequences described for mastadenovirus fibre ...200212018710
development and characterization of bovine x human hybrid cell lines that efficiently support the replication of both wild-type bovine and human adenoviruses and those with e1 deleted.the 293 cell line that was generated by transforming human embryonic kidney cells with human adenovirus type 5 (hav5) early region 1 (e1) sequences is an excellent host for generating and growing hav5 recombinants with e1 deleted, but it does not support the replication of bovine adenovirus type 3 (bav3). madin-darby bovine kidney (mdbk), an established bovine cell line, is an excellent host for growing and plaquing bav3. for the purpose of combining the unique characteristics of these two cell ...200212021321
characterization of bovine adenovirus type 3 e1 proteins and isolation of e1-expressing cell lines.like human adenovirus type 5 (hav5), bovine adenovirus type 3 (bav3) early region 1 (e1) consists of e1a and e1b transcriptional units. in order to characterize bav3 e1 proteins and to isolate a cell line of bovine origin that expresses bav3 e1, polyclonal antibodies specific to e1a, e1b-157r, and e1b-420r were raised in rabbits. bav3 e1a, e1b-157r, and e1b-420r were identified as 40, 17, and 47 kda proteins, and had a half-life of 45-60 min, and 4-6 and 4-6 h, respectively. it appeared that e1a ...200212033770
gene therapy for prostate cancer delivered by ovine adenovirus and mediated by purine nucleoside phosphorylase and fludarabine in mouse models.a gene-directed enzyme pro-drug therapy (gdept) based on purine nucleoside phosphorylase (pnp), that converts the prodrug, fludarabine to 2-fluoroadenine, has been described, but studies are limited compared with other gdepts. we investigated the in vitro and in vivo efficacies of pnp-gdept for treating androgen-independent (ai) prostate cancer. the pnp gene controlled by rous sarcoma virus (rsv) constitutive promoter was delivered using a recombinant ovine adenovirus vector (oadv220) that uses ...200212040457
immunohistopathology of calf pneumonia induced by endobronchial inoculation with bovine adenovirus 3.three 1-week-old and three 3-month-old holstein calves that had received colostrum were inoculated endobronchially with bovine adenovirus 3 (bav-3). the gross and histologic lesions in these six infected calves were localized mainly in the right caudal lobe of the lung and were closely associated with the site of the deposition of the inoculum. the pneumonic lesions were severe necrotizing bronchitis, bronchiolitis, and alveolitis, accompanied by infiltration of inflammatory cells and proliferat ...200212243466
a 72-bp internal deletion in the left inverted terminal repeat of the bovine adenovirus type 3 genome does not affect virus replication.the genome of bovine adenovirus type 3 (bav3) is flanked by 195-base pair (bp) inverted terminal repeats (itr). we isolated a bav3 mutant (bav3c29) having an internal deletion within the left itr. the deletion eliminated 72 bp between nucleotides (nt) 89 and 162, including most of the gc-rich sequences located close to the end of the itr sequences. this deletion did not seem to have any affect on the virus plaque size or morphology and the kinetics of viral replication compared to wild-type (wt) ...200212403925
immuno-gene therapy with adenoviruses expressing fms-like tyrosine kinase 3 ligand and cd40 ligand for mouse hepatoma cells in vivo.introduction of genes encoding immuno-stimulatory cytokines into cancer cells is known to enhance antitumor immunity. cd40 ligand (cd40l, cd154) and fms-like tyrosine kinase 3 ligand (flt3l) are recently identified cytokines, which have been demonstrated to stimulate antitumor immunity in several cancer models. however little is known about antitumor activity of ftl3l and cd40l against hepatocellular carcinoma (hcc). in the present study, we constructed replication-defective adenoviruses express ...200312527933
apoptosis in calf pneumonia induced by endobronchial inoculation with bovine adenovirus type 3 (bav-3).three calves aged 1 week (group 1), three aged 6 weeks (group 2) and three aged 6 weeks (having been pretreated with dexamethasone) (group 3) were infected endobronchially with bovine adenovirus type 3 (bav-3). all calves had received colostrum. the histopathological, immunohistochemical, ultrastructural and tunel features were examined on post-inoculation day (pid) 3, 5 and 7. viral replication and intranuclear inclusions were frequently observed in groups 1 and 3, but not in group 2. the lesio ...200312634091
characterization of the complete genome of the tupaia (tree shrew) adenovirus.the members of the family adenoviridae are widely spread among vertebrate host species and normally cause acute but innocuous infections. special attention is focused on adenoviruses because of their ability to transform host cells, their possible application in vector technology, and their phylogeny. the primary structure of the genome of tupaia adenovirus (tav), which infects tupaia spp. (tree shrew) was determined. tree shrews are taxonomically assumed to be at the base of the phylogenetic tr ...200312634391
bovine adenovirus type 3 pneumonia in dexamethasone-treated calves.the effects of immunosuppression were examined in 1.5-month-old calves that were given dexamethasone (dm) before endobronchial inoculation with bovine adenovirus type 3 (bav-3). immunohistopathologically, severe necrotizing bronchiolitis with eosinophilic and basophilic intranuclear inclusion bodies was observed both in dm-treated 1.5-month-old infected calves and in non-dm-treated 7-day-old infected calves. these inclusion bodies were correlated with the detection of bav-3 antigen and viral par ...200312637751
evaluation of virucidal activity of three commercial disinfectants and formic acid using bovine enterovirus type 1 (ecbo virus), mammalian orthoreovirus type 1 and bovine adenovirus type 1.a modified version of the test method of the comité européen de normalisation (cen) was developed using formic acid and three commercial disinfectants to evaluate virucidal activity against three non-enveloped viruses, bovine enterovirus type 1 (ecbo virus), mammalian orthoreovirus type 1 and bovine adenovirus type 1 (bav 1). determination of the effects of temperature was carried out at 20 and 10 degrees c. all tests with protein load used bovine serum albumin (bsa) and yeast extract. the inves ...200312788019
adenovirus with insertion-mutated e1a selectively propagates in liver cancer cells and destroys tumors in vivo.the adenovirus e1a proteins are involved in the transcriptional activation of viral and cellular genes needed for controlling cell cycle and virus replication. undifferentiated embryonic carcinoma cells have the ability to produce an e1a-like activity that can induce the expression of e1a-targeted adenoviral and cellular genes in the absence of the e1a products. differentiated embryonic carcinoma cells lose the ability to produce the e1a-like activity. in this study, we investigated the e1a-like ...200312810631
adenovirus-mediated heat-activated antisense ku70 expression radiosensitizes tumor cells in vitro and in vivo.ku70 is one component of a protein complex, ku70 and ku80, that functions as a heterodimer to bind dna double-strand breaks and activates dna-dependent protein kinase. our previous study with ku70-/- and ku80-/- mice, and cell lines has shown that ku70- and ku80-deficiency compromises the ability of cells to repair dna double-strand breaks, increases radiosensitivity of cells, and enhances radiation-induced apoptosis. in this study, we examined the feasibility of using adenovirus-mediated, heat- ...200312810658
combination efficacy of doxorubicin and adenoviral methioninase gene therapy with prodrug selenomethionine.we have previously demonstrated an enzyme activation prodrug gene therapy strategy using the methionine alpha,gamma-lyase gene (met) cloned from pseudomonas putida, in combination with selenomethionine (semet) as a prodrug. met gene transfer via a recombinant adenovirus (ad-met) converts the physiologic compound semet to highly toxic methylselenol. in this study, we have developed a combination therapy approach using ad-met/semet gene therapy and doxorubicin (dox). the combination significantly ...200312820369
chemo-radio-gene therapy for colorectal cancer cells using escherichia coli uracil phosphoribosyltransferase gene.5-fluorouracil (5-fu) is one of the most widely used chemotherapeutic agents, and is known to be a radiosensitizer. previously, we reported that adenoviral transduction of the escherichia coli (e. coli) uracil phosphoribosyltransferase (uprt) gene induced marked sensitivity in human colon cancer cells to 5-fu. the aim of the current study was to investigate the efficacy of virally-directed uprt and 5-fu to enhance the radiosensitivity of ht29 human colon cancer cells. cytotoxicity as a result of ...200312820392
gene therapy with gm-csf, interleukin-4 and herpes simplex virus thymidine kinase shows strong antitumor effect on lung cancer.inadequate release of tumor antigens (ta) and a defective antigen presentation by dendritic cells (dc) are the major mechanisms for how tumor cells can escape the host immune surveillance.200312828142
molecular comparison of the fibre proteins of bovine adenovirus subtype a and subtype b.the fibre gene of the bovine adenovirus type 2 (badv2) subtype b was prepared for sequencing by using cloning, sub-cloning and pcr amplification techniques. the nucleotide sequence of the total fibre gene was determined, and it was found to consist of 1,647 nucleotides, coding for a polypeptide of 549 amino acids. the fibre gene regions of badv2 a and b subtypes were aligned. the nucleotide identity of the total fibre gene was found to be 60.5%; however, the homology showed great differences in ...200312837557
crkiii: a novel and biologically distinct member of the crk family of adaptor proteins.as the role for adaptor proteins constantly proliferates, appreciation of their importance has never been higher. the crk family of adaptor proteins is no exception. currently comprising four members, v-crk, crki, crkii and crk-like protein, we have introduced a fifth member, crkiii. cloned by the cort technique, crkiii is identical in sequence to crkii until the second of its two sh3 domains, which is disrupted partway through and results in a nonfunctional domain and a unique c-terminal sequen ...200312894221
new approaches to the prevention and eradication of rabies.despite significant progress in improving the pre- and postexposure prophylaxis of human rabies, the development of better and more cost-effective vaccines and antiviral therapeutics remains a major goal for the treatment of human rabies, the control of animal rabies and particularly for the eradication of rabies virus reservoirs in terrestrial wildlife. in this review, we discuss the structural requirements for an effective rabies vaccine, as well as new strategies currently in use for the deve ...200312903805
molecular anatomy of tupaia (tree shrew) adenovirus genome; evolution of viral genes and viral phylogeny.adenoviruses are globally spread and infect species in all five taxons of vertebrates. outstanding attention is focused on adenoviruses because of their transformation potential, their possible usability as vectors in gene therapy and their applicability in studies dealing with, e.g. cell cycle control, dna replication, transcription, splicing, virus-host interactions, apoptosis, and viral evolution. the accumulation of genetic data provides the basis for the increase of our knowledge about aden ...200312913356
isolation of a bovine adenovirus serotype 10 from a calf in the united states.virus isolated from the lung, liver, kidney, and small intestine of a 3-month-old holstein heifer with a clinical history of pneumonia and lesions in multiple organs was identified as an adenovirus on the basis of morphological and physicochemical characteristics. the adenovirus was determined to be a serotype 10 bovine adenovirus and represents the first reported isolation of this serotype in the united states. inoculation of calves with this isolate resulted in mild to moderate clinical respon ...199912968729
identification of cis-acting sequences required for selective packaging of bovine adenovirus type 3 dna.the assembly of adenovirus particles is a multistep process, in which viral genomic dna is selected and subsequently inserted into preformed empty capsids. the selective encapsidation of the adenovirus genome is directed by cis-acting packaging motifs, termed a repeats due to their at-rich character in dna sequence. a repeats are usually located at the left end of the viral genome. in this report, the construction and analysis of bovine adenovirus type 3 (badv-3) mutants containing deletion muta ...200314573799
construction, rescue, and characterization of vectors derived from ovine atadenovirus.gene transfer vectors derived from ovine atadenovirus type 7 (oadv) can efficiently infect a variety of mammalian cells in vitro and in vivo to deliver and express transgenes. however, early oadv vectors were designed on human mastadenovirus principles prior to the complete characterization of oadv genes and transcripts. the distinctive arrangement of the oadv genome has suggested ways to improve oadv vector design and utility. we therefore developed a cosmid-based approach that allows efficient ...200314581531
altered tropism of recombinant bovine adenovirus type-3 expressing chimeric fiber.recombinant bovine adenovirus-3 (bav-3) has been used as a gene delivery vector for vaccination of calves. however, its usefulness as a vector for non-bovine species is limited due to poor transduction efficiency. to develop bav-3 based vector for non-bovine species, we determined the feasibility of making targeted bav-3 vector by modifying its natural tropism. we constructed a chimeric virus, bav600, in which the knob region of the bav-3 fiber protein was replaced with that from human adenoviru ...200414687941
adenovirus-mediated gene delivery: an overview.adenoviruses, which were first isolated in the 1950s, have been developed as gene-delivery vehicles, or vectors, since the early 1980s. the adenoviruses constitute the adenoviridae family, which is divided into two genera: the aviadenovirus genus infects only birds, whereas the mastadenovirus genus contains viruses that infect a range of mammalian species. human adenoviruses are classified into six subgroups based on the percentage of guanine and cytosine in the dna molecules and the ability to ...200414970581
dna sequencing and analysis of the right-hand part of the genome of the unique bovine adenovirus type 10.the prototype strain of bovine adenovirus (badv) type 10 and four additional isolates that were indistinguishable in serum-neutralization tests have been shown to have remarkable variation in their genome size and restriction maps. in the present study, more than 40 % of the dna sequence of the badv-10 isolate with the longest genome was determined. a biased base composition resulting in low (<41 %) gc content was noticed. analysis of the genes of the dna-binding protein, 100k, 33k, pviii and fi ...200414993642
detection of bovine and porcine adenoviruses for tracing the source of fecal contamination.in this study, a molecular procedure for the detection of adenoviruses of animal origin was developed to evaluate the level of excretion of these viruses by swine and cattle and to design a test to facilitate the tracing of specific sources of environmental viral contamination. two sets of oligonucleotides were designed, one to detect porcine adenoviruses and the other to detect bovine and ovine adenoviruses. the specificity of the assays was assessed in 31 fecal samples and 12 sewage samples th ...200415006765
bovine adenovirus type 3 containing heterologous protein in the c-terminus of minor capsid protein ix.earlier, we detected pix of badv-3 as a 14-kda protein in purified virions. analysis of badv-3 pix using different region antibodies revealed that the n-terminus and central domain of the pix contain immunogenic sites and are not exposed on the surface of badv-3 virion. this suggested that the c-terminus of badv-3 pix (125 amino acid) may be exposed on the virion and may be used as a site for incorporation of heterologous peptides or proteins. we constructed recombinant bav950 containing a small ...200415016551
role of bovine adenovirus-3 33k protein in viral replication.the l6 region of bovine adenovirus type (badv)-3 encodes a nonstructural protein named 33k. to identify and characterize the 33k protein, rabbit polyclonal antiserum was raised against a 33k-gst fusion protein expressed in bacteria. anti-33k serum immunoprecipitated a protein of 42 kda in in vitro translated and transcribed mrna of 33k. however, three proteins of 42, 38, and 33 kda were detected in badv-3 infected cells. to determine the role of this protein in virus replication, a recombinant b ...200415165819
analysis of the first complete genome sequence of an old world monkey adenovirus reveals a lineage distinct from the six human adenovirus species.simian adenovirus 3 (sadv-3) is one of several adenoviruses that were isolated decades ago from old world monkeys. determination of the complete dna sequence of sadv-3 permitted the first full genomic comparison of a monkey adenovirus with adenoviruses of humans (hadvs) and chimpanzees, which are recognized formally as constituting six of the species (hadv-a to hadv-f) within the genus mastadenovirus. the sadv-3 genome is 34 246 bp in size and has a g+c content of 55.3 mol%. it contains all the ...200415448340
characterization and nuclear localization of the fiber protein encoded by the late region 7 of bovine adenovirus type 3.to identify the protein encoded by the l7 region of bovine adenovirus-3 (badv-3), specific antisera were raised by immunizing rabbits with bacterial fusion proteins encoding the n-terminus or c-terminus of the badv-3 fiber protein. immunoprecipitation and western blot analysis confirmed that the fiber is expressed as a 102 kda glycoprotein, which is localized to the nucleus of infected cells. to identify the nuclear localization signals (nls), badv-3 fiber deletion mutants and gfp/beta-galactosi ...200415593420
comparative transduction efficiencies of human and nonhuman adenoviral vectors in human, murine, bovine, and porcine cells in culture.clinical usefulness of human ad serotype 5 (had5) based vectors is limited primarily because of preexisting ad immunity and lack of targeting to specific cell types. alternative vectors based on less prevalent had serotypes as well as nonhuman adenoviruses such as porcine ad serotype 3 (pad3) and bovine ad serotype 3 (bad3) are being developed to overcome these shortcomings. using virus neutralization assay, we examined whether preexisting ad immunity in humans would cross-neutralize pad3 or bad ...200515649439
the coiled-coil domain of the adenovirus type 5 protein ix is dispensable for capsid incorporation and thermostability.the 14.4-kda hexon-associated protein ix (pix) acts as a cement in the capsids of primate adenoviruses and confers a thermostable phenotype. here we show that deletion of amino acids 100 to 114 of adenovirus type 5 pix, which eliminates the conserved coiled-coil domain, impairs its capacity to self-associate. however, pixdelta100-114 is efficiently incorporated into the viral capsid, and the resulting virions are thermostable. deletion of the central alanine-rich domain, as in pixdelta60-72, doe ...200515709043
the effect of paralogous lineages on the application of reconciliation analysis by cophylogeny mapping.paralogy defines similarity caused by duplication rather than common descent and is well known in the case of paralogous gene copies within a single genome. the term is here extended to paralogous lineages of associates within a single host. the phylogenies of four genera within the herpesviridae were reconciled with host phylogenies using cophylogenetic mapping. the observed correspondence for each pair of phylogenies was evaluated through randomization of the viral phylogeny and demonstrated t ...200515805015
evaluation of promoters for foreign gene expression in the e3 region of bovine adenovirus type-3.in order to optimize foreign gene expression in the e3 region of badv-3, we constructed full-length badv-3 genomic dna clones containing a reporter gene (truncated glycoprotein gd of bovine herpesvirus 1, gdt), under the control of exogenous promoters inserted in either direction in the e3 region. irrespective of exogenous transcriptional elements, viable recombinant badv-3 viruses could only be isolated when the gdt expression cassettes were inserted in the e3 region parallel to the direction o ...200515845268
acute respiratory infection with mouse adenovirus type 1.studies of the pathogenesis of adenovirus respiratory disease are limited by the strict species-specificity of the adenoviruses. following intranasal inoculation of adult c57bl/6 mice with mouse adenovirus type 1 (mav-1), we detected mav-1 early region 3 (e3) and hexon gene expression in the lungs at 7 days post-infection (dpi). we detected mav-1 e3 protein in the respiratory epithelium at 7 dpi. we did not detect viral mrna or protein at 14 dpi, but mav-1 dna was detected by pcr at 21 dpi. chem ...200516054189
lh3, a "homologue" of the mastadenoviral e1b 55-kda protein is a structural protein of atadenoviruses.ovine adenovirus serotype 7 (oadv), the prototype atadenovirus, has gene homologues for most mastadenovirus structural proteins but lacks proteins v and ix. instead, oadv has structural proteins of 32 and 42 kda although the gene encoding the latter had not previously been identified. the presently reported studies of oadv virions have now identified a minor structural polypeptide of approximately 40 kda as the product of the l1 52/55-kda gene and, more surprisingly, shown that the 42-kda protei ...200516112161
detection of antibodies against adenovirus protein ix, fiber, and hexon in human sera by immunoblot assay.the 51 serotypes of human adenoviruses (hadvs) of the genus mastadenovirus are classified into the six species hadv-a to hadv-f. for the detection of genus- and species-specific antibodies in human sera an immunoblot assay was developed. the recombinant long fiber of hadv-41[f] (ad41fi) and the native hexon of hadv-5[c] were used as genus-specific antigens. the recombinant capsid protein ix (pix) of hadv-2 (ad2pix[c]) and hadv-41 (ad41pix[f]), the c-terminal pix part of hadv-3 (ad3pixc[b]), and ...200516145087
293t cells expressing simian virus 40 t antigen are semi-permissive to bovine adenovirus type 3 infection.human cells do not normally support productive bovine adenovirus type 3 (badv-3) infection. here, the outcome of badv-3 infection of both 293 cells and 293 cells modified to constitutively express the simian virus 40 (sv-40) t antigen (293t cells) was studied. whereas badv-3 could efficiently infect 293 cells, there was a block in virus dna replication, late-gene expression and virus production. in contrast, replication and efficient virus production could be detected in 293t cells infected with ...200616528030
e1a promoter of bovine adenovirus type 3.conserved motifs of eukaryotic gene promoters, such as tata box and caat box sequences, of e1a of human adenoviruses (e.g human adenovirus 5) lie between the left inverted terminal repeat (itr) and the atg of e1a. however, analysis of the left end of the bovine adenovirus 3 (badv-3) genome revealed that the conserved sequences of the e1a promoter are present only in the itr. as such, the promoter activity of itr was tested in the context of a badv-3 vector or a plasmid-based system. different re ...200617098968
[molecular biology of adenoviral vectors].gene therapy is based on the use of dna as a therapeutic material as an alternative therapeutic tool for treatment of human diseases. all proteins are codified into the dna and several diseases result from the absence or aberrant expression of one or related genes, absence of expression of functional proteins, and alterations for regulation process in transport and degradation mechanisms. in this regard, several diseases could be potentially treated through the expression of the normal form of t ...200617244507
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