RESUMEN
Glucocorticoids regulate the trafficking of cell surface mouse mammary tumor virus (MMTV) glycoproteins in the virus-infected rat hepatoma cell line M1.54. The CR4 rat hepatoma sorting variant, which is derived from M1.54 cells by immunoselection, is uniquely defective in the glucocorticoid-regulated transport of MMTV glycoproteins. Indirect immunofluorescence of fixed permeabilized cells and subcellular fractionation of isolated microsomes revealed that variant CR4 cells retain the MMTV glycoproteins in Golgi-like membranes after glucocorticoid treatment. The variant CR4 phenotype can be complemented by interspecies cell fusions with human HepG2 hepatoma cells and by DNA rescue with genomic fragments isolated from either human or rat hepatoma cells. Transfected wild-type genomic fragments rescue the sorting defect in CR4 at a frequency consistant with a single genetic locus, whereas homologous transfection with CR4 genomic DNA has no effect. Thus, complementation of a rat hepatoma cell-sorting variant supports the existence of a novel protein-trafficking activity encoded by the human or rat genomes that acts in trans in the Golgi to selectively mediate the sorting of cell surface MMTV glycoproteins in glucocorticoid-treated cells.
Asunto(s)
Antígenos Virales de Tumores/genética , Carcinoma Hepatocelular/genética , Glucocorticoides/farmacología , Glicoproteínas/genética , Aparato de Golgi/metabolismo , Virus del Tumor Mamario del Ratón , Proteínas del Envoltorio Viral/genética , Animales , Transporte Biológico , Carcinoma Hepatocelular/patología , ADN/metabolismo , Prueba de Complementación Genética , Variación Genética , Aparato de Golgi/efectos de los fármacos , Humanos , Virus del Tumor Mamario del Ratón/análisis , Fenotipo , Ratas , Células Tumorales CultivadasRESUMEN
Structural proteins designated p10gag, p21gag, p8gag, p3gag, p27gag, and p14gag from the C3H strain of mouse mammary tumor virus (MMTV) were purified by reversed-phase high-pressure liquid chromatography. The N- and C-terminal amino acid sequences and amino acid composition of each protein were determined and compared with the amino acids encoded by the proviral DNA sequences for the MMTV gag gene. The results show that each of the purified proteins is a proteolytic cleavage product derived from the predicted primary translational product of the gag gene (Pr77gag) and that their order in Pr77gag is p10-pp21-p8-p3-n-p27-p14 (where n represents 17 predicted residues that were not identified among the purified proteins). Purified p10gag lacks the initiator methionine and has a myristoyl group attached in amide linkage to the N-terminal glycine residue predicted by the second codon of the gag gene. The cleavage products are contiguous in the sequence of Pr77gag, and the C-terminal residue of p14gag is encoded by the last codon of the gag gene. By analogy with other retrovirus, p14gag is the viral nucleocapsid protein, p10gag is the matrix protein, and p27gag is the capsid protein of mature MMTV. Proteolytic cleavage sites in MMTV Pr77gag bear a striking resemblance to cleavage sites in the gag precursors of D-type retroviruses, suggesting that these viral proteases have similar specificities.
Asunto(s)
Virus del Tumor Mamario del Ratón/análisis , Proteínas de los Retroviridae/análisis , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Cromatografía Líquida de Alta Presión , ADN Viral/genética , Electroforesis en Gel de Poliacrilamida , Productos del Gen gag , Genes Virales , Virus del Tumor Mamario del Ratón/genética , Datos de Secuencia Molecular , Proteínas de los Retroviridae/aislamiento & purificaciónRESUMEN
The chromatin structure of chimeric Moloney murine leukemia viruses (M-MuLVs) containing a glucocorticoid response element (GRE) from mouse mammary tumor virus (MMTV) inserted into the long terminal repeat (LTR) was investigated. Nuclear run-on assays indicated that transcription from the chimeric proviruses was induced 2- to 4-fold by dexamethasone. The wild-type M-MuLV 5' LTR contained a DNase I hypersensitive (HS) site at the TATA sequences, as well as four sites in the enhancer region. The chimeric LTRs contained these sites, as well as three additional sites in the MMTV sequences. Two of the MMTV sites were present in the absence of hormone, while one was hormone-induced. In addition, internal MMTV sequences appeared protected from DNase I digestion in the absence of hormone, suggesting bound protein. Hormone treatment resulted in loss of the DNase I protection.
Asunto(s)
Cromatina/ultraestructura , ADN Viral/análisis , Virus del Tumor Mamario del Ratón/genética , Virus de la Leucemia Murina de Moloney/genética , Provirus/genética , Secuencia de Bases , Quimera , Desoxirribonucleasa I , Dexametasona/farmacología , Glucocorticoides/farmacología , Virus del Tumor Mamario del Ratón/análisis , Virus de la Leucemia Murina de Moloney/análisis , Provirus/análisis , Mapeo Restrictivo , Transcripción Genética/efectos de los fármacos , Cultivo de VirusRESUMEN
The seven mouse mammary tumor virus (MMTV) structural polypeptides (GP52, GP36, P28, P23, P14, P12, and P10) were purified to apparent homogeneity. An improved and reproducible chromatographic method was used to obtain high yields of viral proteins from the same batch of isopycnically purified MMTV. The viral structural proteins were first extracted repeatedly with a high salt and high pH buffer in the presence of 1% Triton X-100. Loss of the minor viral proteins during column purification was minimized by first purifying the smaller molecular weight viral proteins (P14, P12 and P10) by oligo-d(T) column chromatography from the dissociated virions. The other four major viral polypeptides (GP52, GP36, P28 and P23) were then fractionated by affinity and ion-exchange columns. High titer polyclonal antibodies were produced against all of the seven structural proteins except P12 and P10. These antisera were monospecific and showed no cross-reactivity in radioimmunoassay towards other MMTV proteins. With minor modifications, this method has also been applied to purify other structural proteins from several different C-type retroviruses.
Asunto(s)
Virus del Tumor Mamario del Ratón/análisis , Proteínas Virales/aislamiento & purificación , Animales , Cromatografía , Electroforesis en Gel de Poliacrilamida , Sueros Inmunes/inmunología , Ratones , Ratones Endogámicos C3H , Radioinmunoensayo , Proteínas Virales/inmunología , Proteínas Estructurales ViralesRESUMEN
In view of reports that human breast cancer cells secrete growth factors that can replace estradiol in sustaining tumor growth [1], we have investigated whether hormone independent (HI) GR mouse mammary tumors can sustain growth of estrogen-depleted hormone dependent (HD) tumors. HD GR mammary tumor TSl 106 was grafted subcutaneously in the right flank of estrone plus progesterone treated castrated (020 X GR)F1 mice. After 2 weeks the estrone treatment was stopped and the mice received 50, 100 or 150 mg HI GR mammary tumor TSl 104 in the left flank. However, the regression of the HD tumor due to estrone depletion was not prevented or retarded by the HI grafts. In other experiments we investigated integrations of mouse mammary tumor virus (MMTV) proviral DNA in the DNA of GR mammary tumors. We could demonstrate the presence of two cell populations in tumor TSl 96, both HD but differing in MMTV DNA integration events. Our data indicate that exogenous integrations of MMTV proviruses can take place in mouse mammary tumor DNA without loss of hormone dependency of the tumors. Like in GR/Mtv-2+ mice, mammary tumor transplants differing in MMTV proviral integrations are also observed in 020/Mtv-2+ mice.
Asunto(s)
Estrona/farmacología , Neoplasias Mamarias Experimentales/fisiopatología , Neoplasias Hormono-Dependientes/fisiopatología , Progesterona/farmacología , Animales , ADN Viral/análisis , Femenino , Neoplasias Mamarias Experimentales/análisis , Virus del Tumor Mamario del Ratón/análisis , Ratones , Trasplante de Neoplasias , Neoplasias Hormono-Dependientes/análisis , Ovariectomía , Provirus/análisisRESUMEN
The presence of exogenous mouse mammary tumor virus (MMTV) (C3H) DNA sequences in lymphoid tissue (spleen, bone marrow, and thymus) and nonlymphoid tissue (liver and kidney) of BALB/cfC3H female mice was directly assessed by DNA hybridization methods. Lymphoid tissues were found positive for integrated MMTV(C3H) sequences in females as young as 4 weeks. In most samples, the level of splenic MMTV(C3H) infection was low (2 to 5%). Infection remained throughout the life of the animal. The percentage of spleen samples found positive for exogenous viral infection was significantly higher in females bearing mammary tumors, whether virgin or multiparous. Liver and kidney DNAs were negative for exogenous MMTV sequences, suggesting tissue type selectivity in MMTV infection.
Asunto(s)
ADN Viral/análisis , Linfocitos/microbiología , Virus del Tumor Mamario del Ratón/crecimiento & desarrollo , Factores de Edad , Animales , ADN de Neoplasias/análisis , Femenino , Linfocitos/análisis , Tejido Linfoide/microbiología , Neoplasias Mamarias Experimentales/análisis , Virus del Tumor Mamario del Ratón/análisis , Ratones , Ratones Endogámicos , ParidadRESUMEN
From these results we may conclude that estradiol 16 alpha-hydroxylation is highly correlated with tumor incidence, and that the reaction is partly regulated by MMTV and the rest by genetic influences. Elevated hydroxylation appears to be an autosomal dominant trait that is highly specific for estradiol. It is also pertinent that the product of the 16 alpha-hydroxyestrone reaction is a potent estrogen that is capable of binding covalently to amino acids and nucleotides, including the estrogen receptor molecule. The results obtained in these studies establish the usefulness of the mouse model for studying the interrelationship between enhanced 16-hydroxylation of estradiol and the incidence of mammary tumors.
Asunto(s)
Neoplasias de la Mama/análisis , Estradiol/metabolismo , Estriol/análisis , Factores de Edad , Animales , Mama/análisis , Dihidrotestosterona/metabolismo , Estradiol/análogos & derivados , Estradiol/análisis , Femenino , Haplorrinos , Humanos , Hidroxilación , Masculino , Glándulas Mamarias Animales/análisis , Neoplasias Mamarias Experimentales/análisis , Virus del Tumor Mamario del Ratón/análisis , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Pan troglodytes , Pregnenolona/metabolismo , Progesterona/metabolismo , Ratas , Riesgo , Especificidad de la Especie , Esteroide 16-alfa-Hidroxilasa , Testosterona/metabolismoAsunto(s)
Neoplasias de la Mama/patología , Neoplasias Mamarias Experimentales/patología , Lesiones Precancerosas/patología , 9,10-Dimetil-1,2-benzantraceno , Animales , Mama/patología , Transformación Celular Neoplásica , Femenino , Humanos , Glándulas Mamarias Animales/patología , Virus del Tumor Mamario del Ratón/análisis , Metilcolantreno , Ratones , Ratones Endogámicos BALB C , Modelos Biológicos , Mosaicismo , EmbarazoRESUMEN
By electron microscopy and immunocytochemistry we have examined the retroviruses endogenous to AtT20 D16V cells, a cloned line of murine pituitary tumour cells. In addition to the C-type retrovirus particles related to Rauscher murine leukemia virus (MuLV) previously reported to bud from these cells we observed cytoplasmic A-type particles and intracisternal A-type particles (IAP). In the cytoplasm the A-type particles occur in large clusters often associated with sheets of material with a fine structure resembling the shells of the particles. At the plasma membrane individual A-type particles bud to give rise to extracellular virions. The IAP are restricted to the rough endoplasmic reticulum (RER) into which they bud: they are not transported out of the RER to the Golgi apparatus and beyond. We describe a new monoclonal antibody (designated 83E7) which is specific for an epitope of the major core protein (MTVp27) of mouse mammary tumour virus (MMTV). Using immunogold labelling procedures we have specifically labelled both the A-type particles and the associated sheets of material with this antibody. We conclude that the A-type particles and the virions they give rise to are MMTV. The sheets of material must also at least in part be made up of the major core protein of MMTV or its precursor polypeptide. AtT20 cells, therefore, contain endogenous MuLV and MMTV as well as IAP.
Asunto(s)
Genes de Partícula A Intracisternal , Virus del Tumor Mamario del Ratón/ultraestructura , Neoplasias Hipofisarias/microbiología , Proto-Oncogenes , Virus Rauscher/ultraestructura , Animales , Anticuerpos Monoclonales , Especificidad de Anticuerpos , Línea Celular , Invaginaciones Cubiertas de la Membrana Celular/análisis , Técnica del Anticuerpo Fluorescente , Virus del Tumor Mamario del Ratón/análisis , Ratones , Microscopía Electrónica , Neoplasias Hipofisarias/análisis , Neoplasias Hipofisarias/ultraestructura , Virus Rauscher/análisis , Proteínas de los Retroviridae/análisis , Proteínas de los Retroviridae/inmunología , Coloración y Etiquetado , Proteínas del Núcleo Viral/análisis , Proteínas del Núcleo Viral/inmunología , Proteínas del Envoltorio Viral/análisis , Proteínas del Envoltorio Viral/inmunologíaRESUMEN
To determine whether glycopeptides could be released from glycoproteins bound to nitrocellulose, the glycoproteins of murine mammary tumor virus (MuMTV) were radiolabeled by the periodate oxidation/tritiated sodium borohydride reduction technique and separated by gel electrophoresis followed by diffusion transfer. Pronase digestion of nitrocellulose filter strips containing labeled glycoproteins (gp55 or gp34) revealed a rapid release of glycopeptides, i.e., approximately total release within 4 h. The released glycopeptides were similar in size, as determined by molecular sieving chromatography, to glycopeptides obtained by proteolytic digestion of MuMTV glycoproteins from dried gel strips (A. Zilberstein et al., 1980, Cell 21, 417-427) or in solution (M. J. Yagi et al., 1978, Virology 91, 291-304).
Asunto(s)
Glicopéptidos/análisis , Glicoproteínas/análisis , Colodión , Electroforesis en Gel de Poliacrilamida , Virus del Tumor Mamario del Ratón/análisis , Péptido Hidrolasas/farmacología , TritioRESUMEN
Steroid hormones, when complexed to their receptors, recognize and bind specific DNA sequences and subsequently induce increased levels of transcription. The mechanisms of steroid hormone action were analyzed by constructing chimeric DNA molecules from portions of mouse mammary tumor virus envelope and long terminal repeat (LTR) regions ligated to the thymidine kinase (tk) gene of herpes simplex virus. This construction allowed the tk gene to be expressed in a hormone-responsive fashion upon transfection into Ltk- cells. Comparison of transcription data with in vitro binding data showed that hormone-responsive transcription can be directly correlated to the presence of steroid hormone receptor binding sites on the DNA. There are at least two such receptor binding sites in the LTR region, one between -202 and -137 and another between -137 and -50 base pairs from the RNA cap site, as well as a site near the 5' end of the envelope region. These results strengthen the hypothesis that steroid-receptor complexes regulate genes primarily by binding to DNA sites near the promoter region and thereby modulate transcription.
Asunto(s)
ADN Viral/metabolismo , Glucocorticoides/farmacología , Virus del Tumor Mamario del Ratón/análisis , Receptores de Glucocorticoides/metabolismo , Receptores de Esteroides/metabolismo , Transcripción Genética/efectos de los fármacos , Animales , Sitios de Unión , Línea Celular , Quimera , Glucocorticoides/metabolismo , Ratones , Secuencias Repetitivas de Ácidos Nucleicos , Transfección , Triamcinolona Acetonida/metabolismoRESUMEN
A highly leukaemogenic virus isolate (DMBA-LV) endogenous to the CFW/D mouse has been found to contain two viral genomes. One was closely related to the type B milk-borne mouse mammary tumour virus (MMTV) and present in tenfold excess over a type C viral genome which was only partially related to xenotropic and polytropic isolates from the CFW/D mouse as well as to the ecotropic Moloney murine leukaemia virus isolate. The thymic lymphoma cell line that produced DMBA-LV expressed high levels of MMTV viral RNA (35S and the 24S envelope mRNA). Both the virus and the virus-producing cell line expressed multiple species of type C viral RNA. Similar species of type C viral RNA were also associated with non-infectious, non-leukaemogenic viral particles present in both normal lymphoid cells and in a MMTV-free thymic lymphoma cell line established from a second chemical carcinogen-induced tumour.
Asunto(s)
Virus de la Leucemia Murina/aislamiento & purificación , Virus del Tumor Mamario del Ratón/aislamiento & purificación , Animales , Secuencia de Bases , Línea Celular , Genes Virales , Virus de la Leucemia Murina/análisis , Virus de la Leucemia Murina/genética , Linfoma/microbiología , Virus del Tumor Mamario del Ratón/análisis , Virus del Tumor Mamario del Ratón/genética , Ratones , Ratones Endogámicos , Hibridación de Ácido Nucleico , ARN Viral/análisis , ARN Viral/genética , ARN Viral/aislamiento & purificación , Neoplasias del Timo/microbiología , Cultivo de VirusRESUMEN
The mature envelope glycoproteins of mouse mammary tumor virus (gp52 and gp36) were isolated by reversed-phase high-pressure liquid chromatography. The N-terminal amino acid sequence of gp36 was determined for 28 residues. The C-terminal amino acid sequences of gp52 and gp36 were determined by carboxypeptidase digestion. The N-terminal amino acid sequence of gp52 has been reported previously (L. O. Arthur et al., J. Virol. 41:414-422, 1982). These data were aligned with the predicted amino acid sequence of the env gene product obtained by translation of the DNA sequence (S. M. S. Redmond and C. Dickson, Eur. Mol. Biol. Org. J. 2:125-131, 1983). The amino acid sequences of the mature viral proteins were in agreement with the predicted amino acid sequence of the env gene product over the regions of alignment. This alignment showed the sites of proteolytic cleavages of the env gene product leading to the mature viral envelope glycoproteins. The N-terminal amino acid sequence of gp52 starts at residue 99 of the predicted structure indicating proteolytic cleavage of a signal peptide. A dipeptide (Lys-Arg) is excised between the C-terminus of gp52 and the N-terminus of gp36. The C-terminal amino acid sequence of gp36 is identical to the sequence predicted by the codons immediately preceding the termination codon for the env gene product. The data show that there is no proteolytic processing at the C-terminal of the murine mammary tumor virus env gene product and that the env gene coding region extends into the long terminal repeat.
Asunto(s)
Virus del Tumor Mamario del Ratón/análisis , Proteínas Virales/análisis , Secuencia de Aminoácidos , Secuencia de Bases , Genes Virales , Virus del Tumor Mamario del Ratón/genética , Péptidos/metabolismo , Procesamiento Proteico-Postraduccional , Señales de Clasificación de Proteína , Proteínas del Envoltorio Viral , Proteínas Virales/genética , Proteínas Virales/metabolismoRESUMEN
It has recently been shown by mass spectral analysis (Henderson et al., Proc. Natl. Acad. Sci. U.S.A. 80:339-343, 1983) that the p15gag protein of murine leukemia viruses contains a novel post-translational modification, an amino-terminal myristyl (tetradecanoyl) amide. In this report we show that p15gag is the only structural protein to contain this fatty acid. In addition, the gag precursor polyproteins of type B, C, and D retroviruses have been examined for the presence of myristic acid by metabolic labeling and immunoprecipitation studies. In a panel of mammalian type C retroviruses we found that the precursor polyprotein Pr65gag homologs, but not the glycosylated forms (gPr80gag homologs), were specifically labeled after a 5-min incubation of infected cells with [3H]myristic acid. The gag precursor polyprotein was also labeled in mouse mammary tumor virus and in Mason-Pfizer monkey virus, but Pr76gag of Rous sarcoma virus failed to incorporate [3H]myristate. Under similar conditions, [3H]palmitate was not found to be incorporated into any viral gag proteins. Thus, myristylation appears to be a common feature of mammalian type B, C, and D retroviruses but not of avian retroviruses.
Asunto(s)
Ácidos Mirísticos/análisis , Precursores de Proteínas/análisis , Retroviridae/análisis , Proteínas Virales/análisis , Animales , Línea Celular , Productos del Gen gag , Humanos , Virus del Tumor Mamario del Ratón/análisis , Ácidos Mirísticos/metabolismo , Ácido Palmítico , Ácidos Palmíticos/metabolismo , Proteínas Virales/metabolismoRESUMEN
We have detected a mouse mammary tumor virus (MMTV)-specific 1.7-kilobase (kb) polyadenylated RNA in mammary glands of several mouse strains. In BALB/c mice, it is the only MMTV-specific RNA species present. C3H and GR mammary glands and tumors contain, in addition, 3.8- and 7.8-kb MMTV RNAs. Nuclease S1 analysis was performed to map 1.7-kb polyadenylated RNA. It contains predominantly long terminal repeat (LTR) sequences. The 5' end maps approximately 134 nucleotides upstream from the 3' end of the LTR. Colinearity with complete proviral DNA continues to a site about 153 nucleotides downstream from the left (5') LTR. No sequences from the middle part of proviral DNA were found. Colinearity with proviral DNA is resumed 72 nucleotides upstream from the right (3') LTR. The nucleotide sequence in this area is TTCCAGT, which is a splice acceptor consensus sequence. The anatomy of 1.7-kb RNA indicates that it may serve as a messenger for the 36,700-dalton protein encoded by the LTRs of MMTV.
Asunto(s)
Glándulas Mamarias Animales/análisis , Neoplasias Mamarias Experimentales/análisis , Virus del Tumor Mamario del Ratón/análisis , Poli A/análisis , ARN Viral/análisis , ARN/análisis , Animales , Secuencia de Bases , Endonucleasas , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Hibridación de Ácido Nucleico , ARN Mensajero , Secuencias Repetitivas de Ácidos Nucleicos , Endonucleasas Específicas del ADN y ARN con un Solo FilamentoRESUMEN
The major phosphoprotein (p23) of murine mammary tumor virus (MuMTV) was purified for C3H virus, using a combination of alkyl agarose chromatography and gel filtration. Monospecific antiserum was raised against glutaraldehyde crosslinked polymers of purified p23, and the antiserum was then used with iodinated p23 to develop a competition radioimmunoassay. Phosphoprotein p23 was shown to contain both group- and type-specific antigenic determinants in assays using detergent-disrupted virions of MuMTV strains C3H, RIII, GR, and DBA/2 as competing antigens. Tryptic and chymotryptic peptide mapping analysis of p23 polypeptides isolated from MuMTV strains C3H, RIII, GR and DBA/2 showed that the p23s of each virus strain could be clearly distinguished from each other.
Asunto(s)
Antígenos Virales/inmunología , Virus del Tumor Mamario del Ratón/inmunología , Fosfoproteínas/inmunología , Proteínas Virales/inmunología , Epítopos , Virus del Tumor Mamario del Ratón/análisis , Virus del Tumor Mamario del Ratón/clasificación , Fosfoproteínas/aislamiento & purificación , Radioinmunoensayo , Especificidad de la Especie , Proteínas del Núcleo Viral , Proteínas Virales/aislamiento & purificaciónAsunto(s)
Genes Virales , Virus del Tumor Mamario del Ratón/genética , Proteínas Virales/análisis , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Regulación de la Expresión Génica , Productos del Gen gag , Linfocitos/microbiología , Virus del Tumor Mamario del Ratón/análisis , Virus del Tumor Mamario del Ratón/fisiología , Ratones , Morfogénesis , Muridae/microbiología , Precursores de Proteínas/metabolismo , Procesamiento Proteico-Postraduccional , ADN Polimerasa Dirigida por ARN/análisis , Especificidad de la Especie , Proteínas del Envoltorio Viral/análisis , Proteínas Virales/genéticaAsunto(s)
Dexametasona/farmacología , Genes Virales/efectos de los fármacos , Virus del Tumor Mamario del Ratón/genética , Modelos Biológicos , Animales , Carcinoma Hepatocelular/análisis , Carcinoma Hepatocelular/genética , Fenómenos Químicos , Química , Mapeo Cromosómico , Cricetinae , ADN Viral/biosíntesis , Escherichia coli/genética , Genes Bacterianos/efectos de los fármacos , Neoplasias Hepáticas , Linfoma/genética , Neoplasias Mamarias Experimentales/genética , Virus del Tumor Mamario del Ratón/análisis , Virus del Tumor Mamario del Ratón/crecimiento & desarrollo , Ratones , Pentosiltransferasa/genética , Ratas , Receptores de Glucocorticoides/genética , Receptores de Glucocorticoides/fisiología , TransfecciónRESUMEN
We have devised a general infectivity assay for retroviruses. A virus-specific [32P]DNA probe is hybridized in situ to a monolayer culture, and foci of infected cells in the monolayer are detected by exposure of the hybridized culture to X-ray films. The method is quantitative, in that it gives the same titer for Moloney murine leukemia virus as does the standard UV-XC test. The specificity of the assay is indicated by the fact that murine leukemia virus and baboon endogenous virus do not cross hybridize under the conditions used. The assay is completed within 1 to 3 weeks and should be broadly applicable for retroviruses which replicate without altering cellular morphology: its use is demonstrated with mouse mammary tumor virus and the helper virus of the reticuloendotheliosis complex.
Asunto(s)
Hibridación de Ácido Nucleico , ARN Viral/análisis , Retroviridae/análisis , Animales , Gatos , Células Cultivadas , Embrión de Pollo , ADN Viral , Perros , Virus Helper/análisis , Virus del Tumor Mamario del Ratón/análisis , Ratones , Técnicas Microbiológicas , Virus de la Leucemia Murina de Moloney/análisis , Virus de la Reticuloendoteliosis/análisis , Replicación ViralRESUMEN
We have developed a sandwich enzyme immunoassay in order to measure quantitatively mouse mammary tumor virus (MMTV) in mouse milk. In this assay, the antibody-beta-D-galactosidase complex and antibody-bound silicon rubber pieces pieces as solid phase are used. The assay is able to detect 10 ng/ml of MMTV in the milk sample.