RESUMEN

Dendrobium nobile Lindl. is an orcid plant with important medicinal values. This is a colourful houseplant, and also a popular herb in traditional Chinese medicine (TCM). The variants of this plant from different geographic regions might be high, and in this study, we aimed to develop specific sequence characterized amplified region (SCAR) markers for the identification of specific variant of this plant. Different cultivars of D. nobile were collected from nine different places of China, and one cultivar from Myanmar. DNA materials were extracted from the plant samples, random amplified polymorphic DNA (RAPD) were developed, cloned and sequenced for the development of SCAR markers. We have developed four SCAR markers, which are specific to the cultivar from Luzhou China, and clearly distinguishable (genetically) from other cultivars. These SCAR markers are deposited in GenBank (accession number MZ417502, MZ484089, MZ417504 and MZ417505). Four SCAR markers for D. nobile are effective molecular technique to genetically identify the different cultivars or species, and this method is applicable for genetic characterization and identification of other plant species too.

Asunto(s)

Dendrobium , China , ADN , Dendrobium/genética , Marcadores Genéticos/genética , Técnica del ADN Polimorfo Amplificado Aleatorio

RESUMEN

Dendrobium nobile Lindl. is an orcid plant with important medicinal values. This is a colourful houseplant, and also a popular herb in traditional Chinese medicine (TCM). The variants of this plant from different geographic regions might be high, and in this study, we aimed to develop specific sequence characterized amplified region (SCAR) markers for the identification of specific variant of this plant. Different cultivars of D. nobile were collected from nine different places of China, and one cultivar from Myanmar. DNA materials were extracted from the plant samples, random amplified polymorphic DNA (RAPD) were developed, cloned and sequenced for the development of SCAR markers. We have developed four SCAR markers, which are specific to the cultivar from Luzhou China, and clearly distinguishable (genetically) from other cultivars. These SCAR markers are deposited in GenBank (accession number MZ417502, MZ484089, MZ417504 and MZ417505). Four SCAR markers for D. nobile are effective molecular technique to genetically identify the different cultivars or species, and this method is applicable for genetic characterization and identification of other plant species too.

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Dendrobium nobile Lindl. é uma orquídea com importantes valores medicinais. Esta é uma colorida planta doméstica e também uma erva popular na Medicina Tradicional Chinesa (MTC). As variantes desta planta de diferentes regiões geográficas podem ser altas, e neste estudo, nosso objetivo foi desenvolver marcadores de região amplificada de sequência caracterizada (in English, Sequence Characterized Amplified Region (SCAR)) para a identificação de variante específica desta planta. Diferentes cultivares de D. nobile foram coletadas de nove locais diferentes da China e uma cultivar de Mianmar. Materiais de DNA foram extraídos das amostras de plantas, em que a Amplificação Aleatória de DNA Polimórfico (in English, Random Amplified Polymorphism DNA (RAPD)) foi desenvolvida, clonada e sequenciada para o desenvolvimento de marcadores SCAR. Desenvolvemos quatro marcadores SCAR, que são específicos para a cultivar de Luzhou na China e claramente distinguíveis (geneticamente) de outras cultivares. Esses marcadores SCAR estão depositados no GenBank (números de acesso MZ417502, MZ484089, MZ417504 e MZ417505). Quatro marcadores SCAR para D. nobile compreendem técnicas moleculares eficazes para identificar geneticamente as diferentes cultivares ou espécies, e este método é aplicável para caracterização genética e identificação de outras espécies de plantas também.

Asunto(s)

Marcadores Genéticos , Orchidaceae , Dendrobium/genética

RESUMEN

PURPOSE: Detecting different molecular markers in primary tumors and metastases may provide therapeutic information. Here we investigated differences between primary tumors and four metastatic sites of lung adenocarcinoma in the biomarkers' features and discussed potential therapeutic implications. METHODS: A total of 228 patients with metastatic lung adenocarcinoma were analyzed for EGFR, KRAS, BRAF and PIK3CA mutations detected by xTAG liquidchip technology (xTAG-LCT), as well as ERCC1, TYMS, RRM1, TUBB3, STMN1, TOP2A and VEGFR1-3 mRNA expression detected by branched DNA-liquidchip technology (bDNA-LCT). RESULTS: Higher rates of low ERCC1 (35.6 vs. 20.3%, P = 0.0105), RRM1 (23.3 vs. 13.0%, P = 0.0437), STMN1 (72.2 vs. 42.8%, P = 0.0000) and high VEGFR2 (34.4 vs. 18.8%, P = 0.0078) mRNA expression were found in EGFR-mutated tumors, suggesting possible benefit from platinum, gemcitabine, taxanes or VEGFR2 inhibitors. Primary lesions showed low ERCC1 (31.6 vs. 18.5%, P = 0.0271), TYMS (17.6 vs. 7.6%, P = 0.0300), TUBB3 (16.9 vs. 7.6%, P = 0.0415), STMN1 (62.1 vs. 42.9%, P = 0.0065) and high TOP2A (48.7 vs. 33.1%, P = 0.0262) mRNA expression and higher KRAS mutations (25.7 vs. 14.1%, P = 0.0350), suggesting platinum, taxanes, pemetrexed, anti-TOP2A agents and resistant to anti-EGFR therapies. Liver metastases showed absence of low TYMS expression, indicating insensitivity to pemetrexed-based regimen. Pleura metastases harbored higher rates of high VEGFR2 expression (50.0 vs. 19.1%, P = 0.0127). Lymph node metastases presented higher rates of high VEGFR2 expression (37.5 vs. 19.1%, P = 0.0253) and EGFR mutations (59.4 vs. 34.4%, P = 0.0011), suggesting use of anti-VEGFR2 and anti-EGFR therapies. CONCLUSION: Molecular profiling of 228 lung adenocarcinomas determined a significant difference between biomarkers such as EGFR and KRAS subtypes at primary and metastatic sites. Our results serve as a reference for individual treatment based on different potential targets in metastatic lung adenocarcinoma directed by molecular profiling.

Asunto(s)

Adenocarcinoma del Pulmón/secundario , Biomarcadores de Tumor/análisis , Neoplasias Pulmonares/patología , Metástasis de la Neoplasia/patología , Adenocarcinoma del Pulmón/genética , Adulto , Anciano , Femenino , Humanos , Neoplasias Pulmonares/genética , Masculino , Persona de Mediana Edad , Terapia Molecular Dirigida , Metástasis de la Neoplasia/genética

RESUMEN

This study was carried to express the interferon-induced transmembrane protein 3 (IFITM3) in vitro and examine its function in inhibition of avian reovirus (ARV) replication. The recombinant prokaryotic vector expressing yellow-feathered broiler IFITM3 was successfully constructed, and the recombinant protein was expressed in competent Escherichia coli BL21 cells. New Zealand white rabbits were immunized with the purified recombinant protein to prepare a polyclonal antibody, with a titer of 1:128,000. Immunohistochemistry, reverse transcription-PCR, and real-time fluorescence quantitative PCR showed that IFITM3 was distributed in the yellow-feathered broiler immune organs, and the expression of IFITM3 in bursa of Fabricius was more than in spleen and thymus. It was found that in the thymus, spleen and bursa of Fabricius the mRNA expression levels of IFN and IFITM3 were significantly induced after ARV infection. And it was also certified in the chicken embryo fibroblasts (CEFs) which infected with ARV. Then the IFN was added into the cell culture medium before CEFs were infected with ARV. The results indicated that the mRNA of IFITM3 expression was significantly increased and ARV multiplication was significantly inhibited. And when the expression of IFITM3 was knocked down by siRNA-IFITM3, the expression of IFITM3 was significantly reduced, but the ARV multiplication was significantly increased, which indicated that IFITM3 protein could inhibit the ARV replication.

Asunto(s)

Animales , Aves de Corral/virología , Orthoreovirus Aviar , Transporte de Proteínas

RESUMEN

This study was carried to express the interferon-induced transmembrane protein 3 (IFITM3) in vitro and examine its function in inhibition of avian reovirus (ARV) replication. The recombinant prokaryotic vector expressing yellow-feathered broiler IFITM3 was successfully constructed, and the recombinant protein was expressed in competent Escherichia coli BL21 cells. New Zealand white rabbits were immunized with the purified recombinant protein to prepare a polyclonal antibody, with a titer of 1:128,000. Immunohistochemistry, reverse transcription-PCR, and real-time fluorescence quantitative PCR showed that IFITM3 was distributed in the yellow-feathered broiler immune organs, and the expression of IFITM3 in bursa of Fabricius was more than in spleen and thymus. It was found that in the thymus, spleen and bursa of Fabricius the mRNA expression levels of IFN and IFITM3 were significantly induced after ARV infection. And it was also certified in the chicken embryo fibroblasts (CEFs) which infected with ARV. Then the IFN was added into the cell culture medium before CEFs were infected with ARV. The results indicated that the mRNA of IFITM3 expression was significantly increased and ARV multiplication was significantly inhibited. And when the expression of IFITM3 was knocked down by siRNA-IFITM3, the expression of IFITM3 was significantly reduced, but the ARV multiplication was significantly increased, which indicated that IFITM3 protein could inhibit the ARV replication.(AU)

Asunto(s)

Animales , Aves de Corral/virología , Transporte de Proteínas , Orthoreovirus Aviar

RESUMEN

Thirteen new polymorphic microsatellite markers in Fenneropenaeus penicillatus were isolated and characterized. The polymorphism of the thirteen microsatellite markers was tested by 30 individuals from Lianjiang, China. It showed that the number of al-leles per locus ranged from 3 to 6 and the Polymorphism Information Content (PIC) was from 0.324 to 0.706. The observed and expected heterozygosities were 0.3217-0.8023 and 0.1977-0.6783, respectively. Only one loci (LJ-19) deviated significantly from Hardy-Weinberg equilibrium (HWE) (P < 0.00385) after Bonferroni correction, while the other twelve markers were in HWE after Bonferroni correction (P > 0.00385). The thirteen polymorphic microsatellite markers could pro-vide more genetic data for further research on cultivation and recovery of F. penicillatus.

Asunto(s)

Sitios Genéticos , Genética de Población , Repeticiones de Microsatélite , Penaeidae/genética , Alelos , Animales , China , Femenino , Masculino , Polimorfismo Genético

RESUMEN

We investigated the effect of p38MAPK/AP-1 (activator protein-1) signaling on the apoptosis of osteoblasts induced by high glucose. A lentivirus vector of small hairpin RNA (shRNA) targeting p38MAPK was constructed in vitro. Osteoblasts MC3T3-E1 cultured in vitro were treated with vehicle, high glucose, p38MAPK-shRNA transfection, p38MAPK inhibitor, and unrelated shRNA transfection. Apoptosis, protein levels of p38MAPK, and activities of AP-1 in MC3T3-E1 osteoblasts were measured using TUNEL and flow cytometry, Western blot analysis, and an electrophoretic mobility shift assay. Compared with the vehicle group, high glucose induced apoptosis of MC3T3-E1 osteoblasts and activated p38MAPK and AP-1. p38MAPK-shRNA transfection blocked the effect of high glucose stimulation, and the p38MAPK inhibitor showed similar effects as those observed in p38MAPK transfection. Unrelated shRNA had no effect on these changes in MC3T3-E1 osteoblasts induced by high glucose. Therefore, our results suggest that p38MAPK-shRNA reduce apoptosis of MC3T3-E1 osteoblasts induced by high glucose by inhibiting the p38MAPK-AP-1 signaling pathway.

Asunto(s)

Apoptosis/efectos de los fármacos , Glucosa/farmacología , Osteoblastos/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Factor de Transcripción AP-1/metabolismo , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Animales , Western Blotting , Línea Celular , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/farmacología , Citometría de Flujo , Expresión Génica/efectos de los fármacos , Imidazoles/farmacología , Etiquetado Corte-Fin in Situ , Ratones , Osteoblastos/citología , Osteoblastos/metabolismo , Unión Proteica/efectos de los fármacos , Piridinas/farmacología , Interferencia de ARN , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Proteínas Quinasas p38 Activadas por Mitógenos/antagonistas & inhibidores , Proteínas Quinasas p38 Activadas por Mitógenos/genética

RESUMEN

Holothuria leucospilota is a tropical holothurian species that is widely distributed in the tropical and sub-tropical India-Western Pacific Region. Eight polymorphic microsatellite loci were developed from H. leucospilota by using the protocol fast isolation by amplified fragment length polymorphism of sequences containing repeats and tested in 30 individuals from Hainan Island in China. The number of alleles was 2-6 and polymorphism information content ranged from 0.371-0.694. The levels of expected and observed heterozygosities varied from 0.3913-0.6701 and from 0.1154-0.7000, respectively. No significant linkage disequilibrium was detected for any pairwise combination of loci. Only loci YZHS1-42 deviated from Hardy-Weinberg equilibrium. These polymorphic microsatellite loci may be useful for germplasm conservation of H. leucospilota.

Asunto(s)

Sitios Genéticos , Holothuria/genética , Repeticiones de Microsatélite/genética , Polimorfismo Genético , Animales

RESUMEN

The marbled rockfish, Sebastiscus marmoratus, is an important commercially near-shore fish that inhabits the beach rocky bottom from Japan to the South China Sea. Eleven polymorphic microsatellite loci were developed from S. marmoratus and were used to identify polymorphisms in 30 samples from a wild population. The allele locus number ranged from 2 to 7. Polymorphism data content ranged from 0.032 to 0.751. The observed and expected heterozygosity levels were 0.0333-0.9667 and 0.0328-0.7675, respectively. Two loci, Smd1-112 and Smd2-80, deviated from Hardy-Weinberg equilibrium. These polymorphic microsatellite markers will facilitate further studies of genetic diversity and genetic differentiation of S. marmoratus.

Asunto(s)

Peces/genética , Variación Genética , Repeticiones de Microsatélite/genética , Alelos , Animales , China

RESUMEN

The genomic expression profile of the super-hybrid rice Liangyoupeijiu female parent Pei'ai 64S in different tissues at different developmental stages under low temperature, drought, and high temperature stresses were detected using an Affymetrix GeneChip Rice Genome Array to screen upregulated and downregulated genes. In this study, we screened the drought-resistant gene OsRCI2-5, after which a constitutive OsRCI2-5 construct was created and transferred into Nipponbare. After polyethylene glycol-6000 and drought treatment, we found that the OsRCI2-5 gene improved the drought resistance of Nipponbare. Gene expression profiling showed that the OsRCI2-5 gene was expressed in the rice leaves, stems, and flower organs. Subcellular localization revealed that the gene was located in the membranes, and hence, we can deduce that a membrane signal peptide was responsible for signal transduction.

Asunto(s)

Regulación de la Expresión Génica de las Plantas , Oryza/genética , Transducción de Señal/genética , Sequías , Genoma de Planta/genética , Hojas de la Planta/metabolismo , Estrés Fisiológico/genética , Temperatura

RESUMEN

Saccharum spontaneum is the most variable wild relative of sugarcane with potential for use in sugarcane improvement programs. In order to help preserve and exploit this species, 152 accessions from eight major geographical regions in China, including Hainan, Guangdong, Guangxi, Yunnan, Sichuan, Guizhou, Fujian, and Jiangxi provinces, were investigated by analyzing 20 simple sequence repeats (SSRs), including 11 genomic SSRs (gSSRs) and nine SSRs developed from expressed sequence tags (EST-SSRs). A total of 454 alleles were generated by the 20 SSRs, with 295 and 159 alleles detected by gSSRs and EST-SSRs respectively. The Mantel test showed significant correlation between genetic matrixes among the studied accessions revealed by gSSRs versus EST-SSRs, although the average polymorphism of EST-SSRs (17.7) was much lower than that of gSSRs (26.8). Among the eight provinces, collections from Guizhou were the most diverse and those from Guangdong were the most distinct. Clustering analysis and principal component analysis accordantly classified the accessions into four groups, which were "Southwest group", "Hainan group", "Guangdong group", and "Guangxi group", based on the geographical origin of the major accessions in each group, demonstrating that geographical factors play an important role in the pattern of genetic structure of Chinese S. spontaneum. As two (Guizhou and Yunnan) of the three provinces with highest genetic diversity are located in southwest China, we concluded that southwest China is the region with the highest genetic diversity of S. spontaneum.

Asunto(s)

Genes de Plantas , Repeticiones de Microsatélite , Polimorfismo Genético , Saccharum/genética , Alelos , China , Técnicas de Genotipaje , Modelos Genéticos , Filogenia , Filogeografía , Análisis de Componente Principal , Análisis de Secuencia de ADN

RESUMEN

The cytochrome P450c17α gene (CYP17) encodes a key biosynthesis enzyme of estrogen, which is critical in regulating adipogenesis and adipocyte development in humans. We therefore hypothesized that CYP17 is a candidate gene for predicting obesity. In order to test this hypothesis, we performed a family-based association test to investigate the relationship between the CYP17 gene and obesity phenotypes in a large sample comprising 1873 subjects from 405 Caucasian nuclear families of European origin recruited by the Osteoporosis Research Center of Creighton University, USA. Both single SNPs and haplotypes were tested for associations with obesity-related phenotypes, including body mass index (BMI) and fat mass. We identified three SNPs to be significantly associated with BMI, including rs3740397, rs6163, and rs619824. We further characterized the linkage disequilibrium structure for CYP17 and found that the whole CYP17 gene was located in a single-linkage disequilibrium block. This block was observed to be significantly associated with BMI. A major haplotype in this block was significantly associated with both BMI and fat mass. In conclusion, we suggest that the CYP17 gene has an effect on obesity in the Caucasian population. Further independent studies will be needed to confirm our findings.

Asunto(s)

Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Obesidad/enzimología , Obesidad/genética , Esteroide 17-alfa-Hidroxilasa/genética , Población Blanca/genética , Adiposidad/genética , Adulto , Secuencia de Bases , Índice de Masa Corporal , Familia , Femenino , Haplotipos/genética , Humanos , Desequilibrio de Ligamiento/genética , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Polimorfismo de Nucleótido Simple/genética

RESUMEN

Osteoporosis is a heritable disease characterized mainly by low bone mineral density (BMD) and/or osteoporotic fractures (OF). Most genome-wide association studies on osteoporosis have focused on BMD, whereas little effort has been expended to identify genetic variants directly linked to OF. To determine whether BMD-loci are also associated with OF risk, we performed a validation study to examine 23 BMD-loci reported by recent genome-wide association studies for association with hip OF risk. Our sample consisted of 700 elderly Chinese Han subjects, 350 with hip OF and 350 healthy matched controls. We identified four BMD-loci that were significantly associated with hip OF in this Chinese population, including 7q21 (FLJ42280, P = 1.17 × 10(-4) for rs4729260; P = 0.008 for rs7781370), 6p21 (MHC, P = 0.004 for rs3130340), 13q14 (TNFSF11, P = 0.012 for rs9533090; P = 0.018 for rs9594759; P = 0.020 for rs9594738; P = 0.044 for rs9594751), and 18q21 (TNFRSF11A, P = 0.015 for rs884205). The SNP rs4729260 at 7q21 remained significantly associated, even after conservative Bonferroni's correction. Our results further highlight the importance of these loci in the pathogenesis of osteoporosis, and demonstrate that it is feasible and useful to use OF as the direct phenotype to conduct genetic studies, to enhance our understanding of the genetic architecture of osteoporosis.

Asunto(s)

Densidad Ósea/genética , Fracturas de Cadera/genética , Osteoporosis/genética , Fracturas Osteoporóticas/genética , Anciano , Pueblo Asiatico/genética , China/epidemiología , Cromosomas/genética , Femenino , Predisposición Genética a la Enfermedad , Variación Genética , Estudio de Asociación del Genoma Completo , Genotipo , Cadera/patología , Articulación de la Cadera/patología , Humanos , Masculino , Persona de Mediana Edad , Osteoporosis/epidemiología , Osteoporosis/patología , Polimorfismo de Nucleótido Simple

RESUMEN

Mouse PNAS-4 (mPNAS-4) has 96% identity with human PNAS-4 (hPNAS-4) in primary sequence and has been reported to be involved in the apoptotic response to DNA damage. However, there have been no studies reported of the biological functions of mPNAS-4. In studies conducted by our group (unpublished data), it was interesting to note that overexpression of mPNAS-4 promoted apoptotic death in Lewis lung carcinoma cells (LL2) and colon carcinoma cells (CT26) of mice both in vitro and in vivo. In our studies, mPNAS-4 was cloned into the pGEX-6P-1 vector with GST tag at N-terminal in Escherichia coli strain BL21(DE3). The soluble and insoluble expression of recombinant protein mPNAS-4 (rmPNAS-4) was temperature-dependent. The majority of rmPNAS-4 was insoluble at 37 degrees C, while it was almost exclusively expressed in soluble form at 20 degrees C. The soluble rmPNAS-4 was purified by one-step affinity purification, using a glutathione Sepharose 4B column. The rmPNAS-4 protein was further identified by electrospray ionization-mass spectrometry analysis. The search parameters of the parent and fragment mass error tolerance were set at 0.1 and 0.05 kDa, respectively, and the sequence coverage of search result was 28%. The purified rmPNAS-4 was further used as immunogen to raise polyclonal antibodies in New Zealand white rabbit, which were suitable to detect both the recombinant and the endogenous mPNAS-4 in mouse brain tissue and LL2 cells after immunoblotting and/or immunostaining. The purified rmPNAS-4 and our prepared anti-mPNAS-4 polyclonal antibodies may provide useful tools for future biological function studies for mPNAS.

Asunto(s)

Proteínas Reguladoras de la Apoptosis/genética , Apoptosis/fisiología , Células Procariotas/inmunología , Proteínas de Xenopus/genética , Animales , Proteínas Reguladoras de la Apoptosis/inmunología , Proteínas Reguladoras de la Apoptosis/aislamiento & purificación , Western Blotting , ADN Complementario/química , ADN Complementario/genética , Escherichia coli/genética , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Inmunohistoquímica , Ratones , Plásmidos/genética , Conejos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Espectrometría de Masa por Ionización de Electrospray , Proteínas de Xenopus/inmunología , Proteínas de Xenopus/aislamiento & purificación

RESUMEN

Mouse PNAS-4 (mPNAS-4) has 96 percent identity with human PNAS-4 (hPNAS-4) in primary sequence and has been reported to be involved in the apoptotic response to DNA damage. However, there have been no studies reported of the biological functions of mPNAS-4. In studies conducted by our group (unpublished data), it was interesting to note that overexpression of mPNAS-4 promoted apoptotic death in Lewis lung carcinoma cells (LL2) and colon carcinoma cells (CT26) of mice both in vitro and in vivo. In our studies, mPNAS-4 was cloned into the pGEX-6P-1 vector with GST tag at N-terminal in Escherichia coli strain BL21(DE3). The soluble and insoluble expression of recombinant protein mPNAS-4 (rmPNAS-4) was temperature-dependent. The majority of rmPNAS-4 was insoluble at 37°C, while it was almost exclusively expressed in soluble form at 20°C. The soluble rmPNAS-4 was purified by one-step affinity purification, using a glutathione Sepharose 4B column. The rmPNAS-4 protein was further identified by electrospray ionization-mass spectrometry analysis. The search parameters of the parent and fragment mass error tolerance were set at 0.1 and 0.05 kDa, respectively, and the sequence coverage of search result was 28 percent. The purified rmPNAS-4 was further used as immunogen to raise polyclonal antibodies in New Zealand white rabbit, which were suitable to detect both the recombinant and the endogenous mPNAS-4 in mouse brain tissue and LL2 cells after immunoblotting and/or immunostaining. The purified rmPNAS-4 and our prepared anti-mPNAS-4 polyclonal antibodies may provide useful tools for future biological function studies for mPNAS.

Asunto(s)

Animales , Ratones , Conejos , Proteínas Reguladoras de la Apoptosis/genética , Apoptosis/fisiología , Células Procariotas/inmunología , Proteínas de Xenopus/genética , Proteínas Reguladoras de la Apoptosis/inmunología , Proteínas Reguladoras de la Apoptosis/aislamiento & purificación , Western Blotting , ADN Complementario/química , ADN Complementario/genética , Escherichia coli/genética , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Inmunohistoquímica , Plásmidos/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Espectrometría de Masa por Ionización de Electrospray , Proteínas de Xenopus/inmunología , Proteínas de Xenopus/aislamiento & purificación

RESUMEN

In 1992, the era of DNA vaccines began with the report of antibody production upon intradermal injection of mice with a plasmid vector expressing a foreign antigen (1). A rapid succession of subsequent manuscripts showed stimulation of immune responses, including cytolytic T cells, upon inoculation of expression-vectors specific for antigens derived from viruses, bacteria, protozoa and tumor-associated antigens (2-7). Plasmid DNA can be applied through various routes of injection including: intradermal, intramuscular, subcutaneous, intravenous, or directly on mucosal membranes (1,2,8,9). The most commonly used methods of inoculation involve the use of DNA-coated gold beads propelled into the skin by a gene gun or intramuscular inoculation of the vector in saline solution.