RESUMEN
PURPOSE: To compare the range of reach of our newly designed omni-directional ureteroscope (URF-Y0016), compared to the commonly used URF-P6, FlexX2s, and LithoVue™ scopes, in the upper, middle, and lower calyces in an ex-vivo pyelocaliceal model. METHODS: We fabricated a three-dimensional pyelocaliceal model of the upper, middle, and lower pole calyces using urethane and acrylic resin. The inner surface of the dome of each calyx was engraved with reference lines along eight directions, set at 10° of latitude from the top to the base of the dome, and at angles of 0-90°, to precisely determine the range of reach of each scope. The main feature of the URF-Y0016 scope is the omni-directional bending of the tip of the flexible ureteroscope, with the control of these four directions integrated into a handgun-type control unit with a joystick. The range of reach within each calyx was measured by four expert surgeons. RESULTS: The URF-Y0016 scope provided a greater range of reach along all directions in the lower pole calyx compared to URF-P6, FlexX2s, and LithoVue™ scopes (p < 0.001), particularly along the anterior-posterior direction in the lower lobe calyx. However, the URF-Y0016 scope did not influence the improvement of reach range in the upper and middle pole calyx compared to URF-P6, FlexX2s, and LithoVue™ scopes (p = 0.08, p = 0.296). CONCLUSION: The novel design of the URF-Y0016 could improve treatment outcomes for calyceal stones in the lower pole in practice.
Asunto(s)
Pelvis Renal , Ureteroscopios , Diseño de Equipo , Modelos AnatómicosRESUMEN
The existence of tumor-suppressor genes was originally demonstrated by functional complementation through whole-cell and microcell fusion. Transfer of chromosome 11 into a human non-small-cell lung cancer (NSCLC) cell line, A549, suppresses tumorigenicity. Loss of heterozygosity (LOH) on the long arm of chromosome 11 has been reported in NSCLC and other cancers. Several independent studies indicate that multiple tumor-suppressor genes are found in this region, including the gene PPP2R1B at 11q23-24 (ref. 7). Linkage studies of NSCLC are precluded because no hereditary forms are known. We previously identified a region of 700 kb on 11q23.2 that completely suppresses tumorigenicity of A549 human NSCLC cells. Most of this tumor-suppressor activity localizes to a 100-kb segment by functional complementation. Here we report that this region contains a single confirmed gene, TSLC1, whose expression is reduced or absent in A549 and several other NSCLC, hepatocellular carcinoma (HCC) and pancreatic cancer (PaC) cell lines. TSLC1 expression or suppression is correlated with promoter methylation state in these cell lines. Restoration of TSLC1 expression to normal or higher levels suppresses tumor formation by A549 cells in nude mice. Only 2 inactivating mutations of TSLC1 were discovered in 161 tumors and tumor cell lines, both among the 20 primary tumors with LOH for 11q23.2. Promoter methylation was observed in 15 of the other 18 primary NSCLC, HCC and PaC tumors with LOH for 11q23.2. Thus, attenuation of TSLC1 expression occurred in 85% of primary tumors with LOH. Hypermethylation of the TSLC1 promoter would seem to represent the 'second hit' in NSCLC with LOH.
Asunto(s)
Carcinoma de Pulmón de Células no Pequeñas/genética , Genes Supresores de Tumor , Inmunoglobulinas , Neoplasias Pulmonares/genética , Proteínas de la Membrana , Proteínas/genética , Animales , Secuencia de Bases , Molécula 1 de Adhesión Celular , Moléculas de Adhesión Celular , Mapeo Cromosómico , Cromosomas Humanos Par 11 , Cartilla de ADN , ADN Complementario , Ligamiento Genético , Humanos , Pérdida de Heterocigocidad , Ratones , Ratones Desnudos , Datos de Secuencia Molecular , Proteínas Supresoras de TumorRESUMEN
BACKGROUND: The TFII-I is a multifunctional transcriptional factor known to bind specifically to several DNA sequence elements and to mediate growth factor signalling. A microdeletion at the chromosomal location 7q11.23 encoding TFII-I and the related family of transcription factors may result in the onset of Williams-Beuren syndrome, an autosomal dominant genetic disorder characterised by a unique cognitive profile, diabetes, hypertension, anxiety, and craniofacial defects. Hereditary breast and ovarian cancer susceptibility gene product BRCA1 has been shown to serve as a positive regulator of SIRT1 expression by binding to the promoter region of SIRT1, but cross talk between BRCA1 and TFII-I has not been investigated to date. METHODS: A physical interaction between TFII-I and BRCA1 was explored. To determine pathophysiological function of TFII-I, its role as a transcriptional cofactor for BRCA1 was investigated. RESULTS: We found a physical interaction between the carboxyl terminus of TFII-I and the carboxyl terminus of BRCA1, also known as the BRCT domain. Endogenous TFII-I and BRCA1 form a complex in nuclei of intact cells and formation of irradiation-induced nuclear foci was observed. We also showed that the expression of TFII-I stimulates the transcriptional activation function of BRCT by a transient expression assay. The expression of TFII-I also enhanced the transcriptional activation of the SIRT1 promoter mediated by full-length BRCA1. CONCLUSION: These results revealed the intrinsic mechanism that TFII-I may modulate the cellular functions of BRCA1, and provide important implications to understand the development of breast cancer.
Asunto(s)
Proteína BRCA1/fisiología , Factores de Transcripción TFII/fisiología , Animales , Proteína BRCA1/metabolismo , Células COS , Línea Celular Tumoral , Núcleo Celular/metabolismo , Núcleo Celular/patología , Chlorocebus aethiops , Daño del ADN/fisiología , Regulación Neoplásica de la Expresión Génica , Células HeLa , Humanos , Unión Proteica , Sirtuina 1/genética , Sirtuina 1/metabolismo , Transactivadores/metabolismo , Transactivadores/fisiología , Factores de Transcripción TFII/metabolismo , Activación Transcripcional/fisiologíaRESUMEN
BACKGROUND: DBC1/KIAA1967 (deleted in breast cancer 1) is a putative tumour-suppressor gene cloned from a heterozygously deleted region in breast cancer specimens. Caspase-dependent processing of DBC1 promotes apoptosis, and depletion of endogenous DBC1 negatively regulates p53-dependent apoptosis through its specific inhibition of SIRT1. Hereditary breast and ovarian cancer susceptibility gene product BRCA1, by binding to the promoter region of SIRT1, is a positive regulator of SIRT1 expression. METHODS: A physical interaction between DBC1 and BRCA1 was investigated both in vivo and in vitro. To determine the pathophysiological significance of DBC1, its role as a transcriptional factor was studied. RESULTS: We found a physical interaction between the amino terminus of DBC1 and the carboxyl terminus of BRCA1, also known as the BRCT domain. Endogenous DBC1 and BRCA1 form a complex in the nucleus of intact cells, which is exported to the cytoplasm during ultraviolet-induced apoptosis. We also showed that the expression of DBC1 represses the transcriptional activation function of BRCT by a transient expression assay. The expression of DBC1 also inhibits the transactivation of the SIRT1 promoter mediated by full-length BRCA1. CONCLUSION: These results revealed that DBC1 may modulate the cellular functions of BRCA1 and have important implications in the understanding of carcinogenesis in breast tissue.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/fisiología , Proteína BRCA1/metabolismo , Regulación Neoplásica de la Expresión Génica , Activación Transcripcional , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Apoptosis/genética , Proteína BRCA1/química , Proteína BRCA1/fisiología , Células Cultivadas , Células HeLa , Humanos , Unión Proteica , Estructura Terciaria de Proteína/fisiología , Proteínas Represoras/metabolismo , Proteínas Represoras/fisiología , Sirtuina 1/genética , Distribución Tisular , Activación Transcripcional/genéticaRESUMEN
BACKGROUND AND STUDY AIMS: Unsedated transnasal small-caliber esophagogastroduodenoscopy (EGD) has been used to examine the upper gastrointestinal tract with proven feasibility and tolerability. However, a limitation of transnasal EGD is the poor lens-cleansing function of the scope due to the small-caliber water-jet nozzle. Therefore, this trial was designed to evaluate the cleansing effect of oolong tea for transnasal small-caliber EGD. PATIENTS AND METHODS: Oolong tea (O), barley tea (B), and distilled water (W) were prepared as washing solutions for endoscopic lenses. Study I: after the lenses were soiled by lard oil, they were washed with one of the three washing solutions, and the image quality of photographs was judged. Study II: 982 patients who were due to undergo transnasal EGD were enrolled and randomly assigned to the O-, B-, or W-groups. The level of lens cleansing, the overall time required for endoscopy, and the volume of washing solution used were measured. RESULTS: Study I: the image quality of photographs taken with lenses washed with oolong tea was significantly superior to that associated with other solutions. Study II: the level of lens cleansing in the O-group was significantly superior to that of the B- and W-groups ( P < 0.001). The volume of solution used for lens cleansing in the O-group was significantly smaller than that in the W-group ( P < 0.05). Endoscopic examination times in the O-group were shorter than those in the B- and W-groups ( P < 0.05). CONCLUSIONS: In transnasal small-caliber EGD, oolong tea instead of water as a washing solution for endoscopic lens cleansing is useful to maintain good visibility.
Asunto(s)
Bebidas , Detergentes/farmacología , Desinfección/métodos , Endoscopios Gastrointestinales , Lentes , Té , Femenino , Estudios de Seguimiento , Humanos , Masculino , Persona de Mediana Edad , Nariz , Estudios ProspectivosRESUMEN
Reg I (regenerating gene product I) is a growth factor that plays a central role in the generation and regeneration of the gastric mucosal architecture. On the other hand, mouse Reg I mRNA is expressed at the highest levels in the small intestine among the gastrointestinal tissues. In the current study, with the aim to clarify the role of Reg I protein in the small intestine, the temporal and spatial pattern of Reg I expression and the phenotype of Reg I-knockout mice in the tissue were examined. In the wild-type mice, immunohistochemistry localized Reg I protein expression in absorptive cells located in the lower half of the intestinal villi. Reg I expression was undetectable until embryonic day 13 (E13), when the fetal intestine still lacks villous structure; however, it dramatically increased at E17 along with the formation and maturation of the fetal intestinal villi. In the small intestine of the adult Reg I-knockout mice, less densely packed, round-shaped aberrant morphology of the absorptive cells was observed light microscopically, and electron microscopical examination revealed a strikingly loose connection of these cells to the basement membrane. Antiproliferating cell nuclear antigen staining and anti-Ki67 staining demonstrated the marked decrease in the number of proliferating cells in the small intestinal mucosa of the knockout mice. The cell migration speed visualized by one shot labeling of 5-bromodeoxyuridine was significantly slower in the knockout mice. These phenotypes of Reg I-knockout mice emerged, in accordance with the temporal pattern of Reg I expression described above, from E17. Reg I was considered to be a regulator of cell growth that is required to generate and maintain the villous structure of the small intestine.
Asunto(s)
Proliferación Celular , Mucosa Intestinal/citología , Intestino Delgado/citología , Litostatina/fisiología , Microvellosidades/ultraestructura , Animales , Procesos de Crecimiento Celular , Movimiento Celular , Femenino , Regulación del Desarrollo de la Expresión Génica , Mucosa Intestinal/ultraestructura , Intestino Delgado/ultraestructura , Litostatina/genética , Masculino , Ratones , Ratones Endogámicos ICR , Ratones Noqueados , Fenotipo , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The transportation of poultry and related products for international trade contributes to transboundary pathogen spread and disease outbreaks worldwide. To prevent pathogen incursion through poultry products, many countries have regulations about animal health and poultry product quarantine. However, in Japan, animal products have been illegally introduced into the country in baggage and confiscated at the airport. Lately, the number of illegally imported poultry and the incursion risk of transboundary pathogens through poultry products have been increasing. In this study, we isolated avian influenza viruses (AIVs) from raw poultry products illegally imported to Japan by international passengers. Highly (H5N1 and H5N6) and low (H9N2 and H1N2) pathogenic AIVs were isolated from raw chicken and duck products carried by flight passengers. H5 and H9 isolates were phylogenetically closely related to viruses isolated from poultry in China, and haemagglutinin genes of H5N1 and H5N6 isolates belonged to clades 2.3.2.1c and 2.3.4.4, respectively. Experimental infections of H5 and H9 isolates in chickens and ducks demonstrated pathogenicity and tissue tropism to skeletal muscles. To prevent virus incursion by poultry products, it is important to encourage the phased cleaning based on the disease control and eradication and promote the reduction in contamination risk in animal products.
Asunto(s)
Aeropuertos , Comercio , Subtipo H1N2 del Virus de la Influenza A/aislamiento & purificación , Subtipo H5N1 del Virus de la Influenza A/aislamiento & purificación , Subtipo H9N2 del Virus de la Influenza A/aislamiento & purificación , Gripe Aviar/virología , Productos Avícolas/virología , Viaje , Animales , Antígenos Virales/inmunología , Pollos/virología , China/epidemiología , Brotes de Enfermedades/prevención & control , Brotes de Enfermedades/veterinaria , Patos/virología , Microbiología de Alimentos , Subtipo H1N2 del Virus de la Influenza A/genética , Subtipo H1N2 del Virus de la Influenza A/inmunología , Subtipo H5N1 del Virus de la Influenza A/genética , Subtipo H5N1 del Virus de la Influenza A/inmunología , Subtipo H9N2 del Virus de la Influenza A/genética , Subtipo H9N2 del Virus de la Influenza A/inmunología , Gripe Aviar/epidemiología , Japón , Carne/virología , Filogenia , Aves de Corral/virología , Enfermedades de las Aves de Corral/epidemiología , ARN Viral/genéticaRESUMEN
At variance with the earlier belief that mitochondrial genomes are represented by circular DNA molecules, a large number of organisms have been found to carry linear mitochondrial DNA. Studies of linear mitochondrial genomes might provide a novel view on the evolutionary history of organelle genomes and contribute to delineating mechanisms of maintenance and functioning of telomeres. Because linear mitochondrial DNA is present in a number of human pathogens, its replication mechanisms might become a target for drugs that would not interfere with replication of human circular mitochondrial DNA.
Asunto(s)
ADN Mitocondrial , Animales , Evolución Biológica , Replicación del ADN , Humanos , TelómeroRESUMEN
The mitochondrial deoxyribonucleic acid (mtDNA) from a petite-negative yeast, Hansenula mrakii, was studied. A linear restriction map was constructed with 11 restriction enzymes. The linearity of the genome was confirmed by direct end labeling of the molecule, followed by restriction analysis. The molecular weight of the DNA was found to be 55,000 base pairs. This is the first linear mtDNA found in yeast species. Using specific gene probes obtained from Saccharomyces cerevisiae mtDNA, we have constructed a gene map of H. mrakii mtDNA. The arrangement of genes in this linear genome was very different from the circular mtDNA of other known yeasts.
Asunto(s)
ADN de Hongos/genética , ADN Mitocondrial/genética , Pichia/genética , Saccharomycetales/genética , Mapeo Cromosómico , ADN de Hongos/aislamiento & purificación , ADN Mitocondrial/aislamiento & purificación , Peso Molecular , Especificidad de la EspecieRESUMEN
In yeast mitochondria, most of the isoaccepting species of tyrosyl tRNA are coded by a mitochondrial gene, tyrA. A particular isoaccepting species is coded by a second mitochondrial gene, tyrB. This gene is not expressed in certain strains of yeast which show no deficient phenotype. Genetic crosses between strains expressing or not expressing the tyrB gene demonstrate that expression is controlled by specific nuclear genes and that a mutation of the tyrA gene can be bypassed when the tyrB gene is operative.
Asunto(s)
Regulación de la Expresión Génica , Aminoacil-ARN de Transferencia/genética , Saccharomyces cerevisiae/genética , Núcleo Celular/análisis , Mitocondrias/análisisRESUMEN
The terminal structure of the linear mitochondrial DNA (mtDNA) from three yeast species has been examined. By enzymatic digestion, alkali denaturation, and sequencing of cloned termini, it was shown that in Pichia pijperi and P. jadinii, both termini of the linear mtDNA were made of a single-stranded loop covalently joining the two strands, as in the case of vaccinia virus DNA. The left and right loop sequences were in either of two orientations, suggesting the existence of a flip-flop inversion mechanism. Contiguous to the terminal loops, inverted terminal repeats were present. The mtDNA from Williopsis mrakii seems to have an analogous structure, although terminal loops could not be directly demonstrated. Electron microscopy revealed the presence, among linear molecules, of a small number of circular DNAs, mostly of monomer length. Linear and circular models of replication are considered, and possible conversion mechanisms between linear and circular forms are discussed. A flip-flop inversion mechanism between the inverted repeat sequences within a circular intermediate may be involved in the generation of the linear form of mtDNA.
Asunto(s)
ADN de Hongos/ultraestructura , ADN Mitocondrial/ultraestructura , Levaduras/genética , Secuencia de Bases , Clonación Molecular , Replicación del ADN , ADN Circular/genética , ADN Circular/ultraestructura , ADN de Hongos/química , ADN Mitocondrial/química , Exodesoxirribonucleasas/farmacología , Concentración de Iones de Hidrógeno , Microscopía Electrónica , Datos de Secuencia Molecular , Desnaturalización de Ácido Nucleico , Secuencias Repetitivas de Ácidos NucleicosRESUMEN
The RAG1 gene of Kluyveromyces lactis encodes a low-affinity glucose/fructose transporter. Its transcription is induced by glucose, fructose, and several other sugars. The RAG4, RAG5, and RAG8 genes are trans-acting genes controlling the expression of the RAG1 gene. We report here the characterization of one of these genes, RAG5. The nucleotide sequence of the cloned RAG5 gene indicated that it encodes a protein that is homologous to hexokinases of Saccharomyces cerevisiae. rag5 mutants showed no detectable hexokinase or glucokinase activity, suggesting that the sugar kinase activity encoded by this gene is the only hexokinase in K. lactis. Both high- and low-affinity transport systems of glucose were affected in rag5 mutants. The defect of the low-affinity component was found to be due to a block of transcription of the RAG1 gene by the hexokinase mutation. In vivo complementation of the rag5 mutation by the HXK2 gene of S. cerevisiae and complementation of hxk1 hxk2 mutations of S. cerevisiae by the RAG5 gene showed that RAG5 and HXK2 were equivalent for sugar-phosphorylating activity but that RAG5 could not restore glucose repression in the S. cerevisiae hexokinase mutants.
Asunto(s)
Genes Fúngicos , Hexoquinasa/genética , Kluyveromyces/genética , Proteínas de Transporte de Monosacáridos/genética , Transcripción Genética , Secuencia de Aminoácidos , Northern Blotting , Clonación Molecular , Regulación Fúngica de la Expresión Génica/genética , Prueba de Complementación Genética , Hexoquinasa/metabolismo , Kluyveromyces/enzimología , Kluyveromyces/crecimiento & desarrollo , Datos de Secuencia Molecular , Proteínas de Transporte de Monosacáridos/metabolismo , Fenotipo , Fosforilación , Mapeo Restrictivo , Homología de Secuencia de AminoácidoRESUMEN
In most yeast species, the mitochondrial DNA (mtDNA) has been reported to be a circular molecule. However, two cases of linear mtDNA with specific termini have previously been described. We examined the frequency of occurrence of linear forms of mtDNA among yeasts by pulsed-field gel electrophoresis. Among the 58 species from the genera Pichia and Williopsis that we examined, linear mtDNA was found with unexpectedly high frequency. Thirteen species contained a linear mtDNA, as confirmed by restriction mapping, and labeling, and electron microscopy. The mtDNAs from Pichia pijperi, Williopsis mrakii, and P. jadinii were studied in detail. In each case, the left and right terminal fragments shared homologous sequences. Between the terminal repeats, the order of mitochondrial genes was the same in all of the linear mtDNAs examined, despite a large variation of the genome size. This constancy of gene order is in contrast with the great variation of gene arrangement in circular mitochondrial genomes of yeasts. The coding sequences determined on several genes were highly homologous to those of the circular mtDNAs, suggesting that these two forms of mtDNA are not of distant origins.
Asunto(s)
ADN de Hongos/química , ADN Mitocondrial/química , Levaduras/genética , Secuencia de Bases , Clasificación , Electroforesis en Gel de Campo Pulsado , Genes Fúngicos , Ligamiento Genético , Microscopía Electrónica , Datos de Secuencia Molecular , Mapeo Restrictivo , Homología de Secuencia de Ácido NucleicoRESUMEN
BACKGROUND: Pit pattern diagnosis is important for endoscopic detection of dysplastic Barrett's lesions, though using magnification endoscopy can be difficult and laborious. We investigated the usefulness of a modified crystal violet chromoendoscopy procedure and utilised a new pit pattern classification for diagnosis of dysplastic Barrett's lesions. METHODS: A total of 1,030 patients suspected of having a columnar lined oesophagus were examined, of whom 816 demonstrated a crystal violet-stained columnar lined oesophagus. The early group of patients underwent 0.05% crystal violet chromoendoscopy, while the later group was examined using 0.03% crystal violet with 3.0% acetate. A targeted biopsy of the columnar lined oesophagus was performed using crystal violet staining after making a diagnosis of closed or open type pit pattern with a newly proposed system of classification. The relationship between type of pit pattern and histologically identified dysplastic Barrett's lesions was evaluated. RESULTS: Dysplastic Barrett's lesions were identified in biopsy samples with an open type pit pattern with a sensitivity of 96.0%. Further, Barrett's mucosa with the intestinal predominant mucin phenotype was closely associated with the open type pit pattern (sensitivity 81.9%, specificity 95.6%). CONCLUSIONS: The new pit pattern classification for diagnosis of Barrett's mucosa was found to be useful for identification of cases with dysplastic lesions and possible malignant potential using a crystal violet chromoendoscopic procedure.
Asunto(s)
Antiinfecciosos Locales , Esófago de Barrett/patología , Endoscopía del Sistema Digestivo/métodos , Violeta de Genciana , Acetatos , Adulto , Anciano , Anciano de 80 o más Años , Esófago de Barrett/clasificación , Biopsia , Esófago/patología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Membrana Mucosa/patologíaRESUMEN
Loss of heterozygosity on chromosome 10p was observed frequently in human prostate cancers. Studies have demonstrated that the introduction of the short arm of human chromosome 10 into a human prostate cancer cell line, PPC-1, by microcell-mediated chromosome transfer (MMCT), suppressed the malignant phenotype, suggesting the presence of a prostate tumor suppressor gene(s) within a region of 17 cM at distal 10p. To narrow down the candidate region harboring the tumor suppressor gene, a series of 10p fragments were transferred into PPC-1 cells by MMCT using a panel of hamster-human hybrid cells containing various portions of 10p. Four of the six hybrid cells obtained showed decreased tumorigenicity when injected subcutaneously into athymic nude mice. Tumors developed only at six of 40 injection sites for these four hybrid cells. In contrast, the other two hybrid cells, as well as parental PPC-1 cells, were judged to be fully tumorigenic because tumors appeared at a total 26 of 32 sites for the two hybrid cells and 15 of 16 sites for PPC-1. Allelotyping of 10p combined with fluorescence in situ hybridization in these hybrid cells suggested that a prostate tumor suppressor gene was located within a fragment of approximately 1.2 Mb flanked by D10S1172 and D10S226 on 10p15.1.
Asunto(s)
Cromosomas Humanos Par 10 , Genes Supresores de Tumor , Neoplasias de la Próstata/genética , Adenocarcinoma/genética , Adenocarcinoma/patología , Animales , Pruebas de Carcinogenicidad , Cricetinae , Técnicas de Transferencia de Gen , Marcadores Genéticos , Humanos , Células Híbridas , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Neoplasias de la Próstata/patología , Lugares Marcados de Secuencia , Células Tumorales CultivadasRESUMEN
We have recently identified the TSLC1 gene as a novel tumor suppressor in human non-small cell lung cancers. TSLC1 encodes a membrane glycoprotein with an extracellular domain homologous to those of immunoglobulin superfamily proteins. Truncation of TSLC1 in the cytoplasmic domain in a primary human tumor suggests that this domain is important for tumor suppressor activity. Here, we report the isolation of two TSLC1-like genes, TSLL1 and TSLL2, based on their structural homology with the sequences corresponding to the cytoplasmic domain of TSLC1. Significant similarity was also observed in the extracellular domain as well as in the overall gene structure, indicating that these three genes form a unique subfamily (the TSLC1-gene family) in the immunoglobulin superfamily genes. In contrast to the ubiquitous expression of TSLC1, TSLL1 is expressed exclusively in adult and fetal human brain, while TSLL2 is expressed in several specific tissues including prostate, brain, kidney and some other organs. Expression of TSLL1 and TSLL2 was lost or markedly reduced in many human glioma cell lines or some prostate cancer cell lines, suggesting that loss of expression of these genes might be involved in some human cancers.
Asunto(s)
Inmunoglobulinas , Proteínas de la Membrana , Biosíntesis de Proteínas , Proteínas/genética , Adulto , Secuencia de Aminoácidos , Secuencia de Bases , Northern Blotting , Encéfalo/embriología , Carcinoma de Pulmón de Células no Pequeñas/genética , Molécula 1 de Adhesión Celular , Moléculas de Adhesión Celular , Membrana Celular/metabolismo , Mapeo Cromosómico , Clonación Molecular , ADN Complementario/metabolismo , Exones , Humanos , Hibridación Fluorescente in Situ , Intrones , Neoplasias Pulmonares/genética , Modelos Genéticos , Datos de Secuencia Molecular , Familia de Multigenes , Estructura Terciaria de Proteína , Proteínas/química , Homología de Secuencia de Aminoácido , Distribución Tisular , Células Tumorales Cultivadas , Proteínas Supresoras de TumorRESUMEN
Fifteen tRNA-encoding genes were mapped on the linear mitochondrial (mt) DNA of the yeast, Candida parapsilosis, and their sequences determined. The gene order and gene sequences indicate that the mt genome of this yeast belongs to a group clearly different from the already known group of linear mt DNAs of yeast.
Asunto(s)
Candida/genética , ADN Mitocondrial/química , Genes Fúngicos/genética , ARN de Transferencia/genética , Secuencia de Bases , Mapeo Cromosómico , ADN de Hongos/química , ADN de Hongos/genética , ADN Mitocondrial/genética , Datos de Secuencia Molecular , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
The aminoglycoside 3'-phosphotransferase type I (APHI)-coding gene of the bacterial transposon Tn903 confers resistance to kanamycin on bacteria and resistance to geneticin (G418) on many eukaryotes. We developed an APHI fusion system that can be used in the study of gene expression in these organisms, particularly in yeasts. The first 19 codons of the KmR (APHI) gene can be deleted, and replaced by other genes in a continuous reading frame, without loss of APH activity. Examples of vector constructions are given which are adapted to the yeast Kluyveromyces lactis transformation system. Their derivatives containing the 2 mu origin of replication can also be used in Saccharomyces cerevisiae.
Asunto(s)
Clonación Molecular , Elementos Transponibles de ADN , Genes Fúngicos , Genes , Resistencia a la Kanamicina/genética , Kluyveromyces/genética , Fosfotransferasas/genética , Saccharomyces cerevisiae/genética , Saccharomycetales/genética , Secuencia de Bases , Kanamicina Quinasa , Kluyveromyces/enzimología , Datos de Secuencia Molecular , Plásmidos , Regiones Promotoras Genéticas , Saccharomyces cerevisiae/enzimologíaRESUMEN
The efficiency of secreted production of mammalian proteins from yeasts remains unpredictably variable, depending on each protein. On the hypothesis that the control of protein conformation during protein translocation is the bottleneck in many cases, we examined the effects of an increased dosage of the genes coding for protein disulfide isomerase and of polyubiquitin on the secretion of two human proteins, serumalbumin and interleukin 1 beta. The yeast Kluyveromyces lactis was used as a production host. Duplication of either one of these genes had a strong stimulating effect on the production of the highly disulfide-bonded serumalbumin, but not of interleukin 1 beta.