Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 23
Filter
1.
Chinese Journal of Biotechnology ; (12): 1363-1373, 2023.
Article in Chinese | WPRIM | ID: wpr-981143

ABSTRACT

The CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR associated proteins) system is an adaptive immune system of bacteria and archaea against phages, plasmids and other exogenous genetic materials. The system uses a special RNA (CRISPR RNA, crRNA) guided endonuclease to cut the exogenous genetic materials complementary to crRNA, thus blocking the infection of exogenous nucleic acid. According to the composition of the effector complex, CRISPR-Cas system can be divided into two categories: class 1 (including type Ⅰ, Ⅳ, and Ⅲ) and class 2 (including type Ⅱ, Ⅴ, and Ⅵ). Several CRISPR-Cas systems have been found to have very strong ability to specifically target RNA editing, such as type Ⅵ CRISPR-Cas13 system and type Ⅲ CRISPR-Cas7-11 system. Recently, several systems have been widely used in the field of RNA editing, making them a powerful tool for gene editing. Understanding the composition, structure, molecular mechanism and potential application of RNA-targeting CRISPR-Cas systems will facilitate the mechanistic research of this system and provide new ideas for developing gene editing tools.


Subject(s)
CRISPR-Cas Systems/genetics , RNA/genetics , Bacteria/genetics , Gene Editing , Archaea
2.
Journal of Peking University(Health Sciences) ; (6): 392-399, 2023.
Article in Chinese | WPRIM | ID: wpr-986867

ABSTRACT

OBJECTIVE@#To investigate the effect of titanium dioxide nanoparticles (TiO2 NPs) on the expression profile of circular ribonucleic acid (circRNA) in human hepatocytes through in vitro cell experiments, and to attempt to understand the potential mechanism of hepatotoxicity through bioinformatics analysis.@*METHODS@#TiO2 NPs were characterized from the aspects of particle size, shape and agglomeration state. The cell counting kit-8 (CCK8) was used to detect the cytotoxicity of TiO2 NPs against human hepatocellular carcinoma cells (HepG2) after exposure to 0, 1.56, 3.13, 6.25, 12.5, 25, 50, 100, and 200 mg/L TiO2 NPs for 24 h or 48 h. The cells were treated at doses of 0 mg/L TiO2 NPs (control group) and 100 mg/L TiO2 NPs (treatment group), and collected after exposure for 48 h, and then RNA from the extracted cell samples was collected and sequenced. The differential circRNAs between the control and the TiO2 NPs treatment groups were screened, and then the enrichment pathway of the differential circRNA target gene was analyzed by multivariate statistics. According to the sequencing results, significantly altered genes and important genes in the significant enrichment pathways were screened, and real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) was performed to verify the results.@*RESULTS@#TiO2 NPs were spherical anatase with a hydrated particle size of (323.50±85.44) nm and a Zeta potential of (-21.00±0.72) mV in a serum-free medium. The results of the CCK8 cytotoxicity assay showed that with the increase of TiO2 NPs concentration, cell viability gradually decreased. A total of 11 478 circRNAs were found by RNA sequencing. Compared with the control groups, TiO2 NPs treatment groups (100 mg/L) had a total of 89 differential circRNAs, of which 59 were up-regulated and 30 were down-regulated. Analysis of the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway showed that the targeted genes of differential circRNAs were mainly enriched in fatty acid degradation, Fanconi anemia pathway, and fatty acid metabolism. The expression levels of circRNA.6730, circRNA.3650 and circRNA.4321 were significantly different between the TiO2 NPs treatment group and the control group, which were consistent with the sequencing results.@*CONCLUSION@#TiO2 NPs can induce changes in circRNA expression profile, and epigenetics may play an important role in the mechanism of hepatotoxicity.


Subject(s)
Humans , RNA/genetics , RNA, Circular/genetics , Carcinoma, Hepatocellular/genetics , Liver Neoplasms/genetics , Titanium , Nanoparticles , Chemical and Drug Induced Liver Injury , Fatty Acids
3.
Bol. micol. (Valparaiso En linea) ; 36(1): 12-16, jun. 2021. ilus
Article in Spanish | LILACS | ID: biblio-1380384

ABSTRACT

Se describe hasta la fecha de hoy, 4 de julio del 2021, la evidencia existente sobre la variante Delta del SARS-CoV-2, su impacto en la trasmisión, en la severidad de la infección y su probable evasión a la respuesta inmune. (AU)


As of today, July 4, 2021, the existing evidence on the Delta variant of SARS-CoV-2, its impact on transmission, on the severity of the infection and its probable evasion of the immune response is described. (AU)


Subject(s)
Humans , RNA/genetics , COVID-19/genetics , Mutation/genetics , Chile/epidemiology , Mass Vaccination , Immune Evasion , COVID-19/transmission , COVID-19/epidemiology
4.
Chinese Journal of Contemporary Pediatrics ; (12): 730-734, 2021.
Article in Chinese | WPRIM | ID: wpr-888473

ABSTRACT

OBJECTIVE@#To determine the association of circular RNA (circRNA) and circRNA-microRNA (miRNA) network regulation with brain injury induced by inflammation in preterm mice.@*METHODS@#Pregnant mice were treated with intraperitoneally injected lipopolysaccharide to establish a preterm mouse model of brain injury induced by inflammation (inflammation preterm group with 3 mice). Preterm mice born to normal pregnant mice by cesarean section were selected as controls (non-inflammation preterm group with 3 mice). The gene microarray technique was used to screen out the circRNAs associated with brain injury in preterm mice. The miRNA target prediction software was used to predict the binding sites between circRNAs and miRNAs and analyze the regulatory mechanism.@*RESULTS@#A total of 365 differentially expressed circRNAs were screened out between the inflammation preterm and non-inflammation preterm groups (fold change > 1.5, @*CONCLUSIONS@#Inflammation induces a significant change in the expression profile of circRNAs in the brain tissue of mice, and the change in the expression of circRNAs plays an important role in brain injury induced by inflammation and subsequent brain development in preterm mice.


Subject(s)
Animals , Female , Mice , Pregnancy , Brain Injuries , Cesarean Section , Gene Expression Profiling , Inflammation/genetics , MicroRNAs/genetics , RNA/genetics , RNA, Circular
5.
Chinese Medical Journal ; (24): 2573-2582, 2021.
Article in English | WPRIM | ID: wpr-921184

ABSTRACT

BACKGROUND@#Circular RNA (circRNA) is a type of closed circular noncoding RNA (ncRNA), mostly formed by back-splicing or alternative splicing of pre-messenger RNA (mRNA). The aim of this study was to explore the expression profile of circRNA in peripheral blood mononuclear cells (PBMCs) of patients with ankylosing spondylitis (AS) and discover potential molecular markers of AS.@*METHODS@#The circRNA microarray technology was used to detect the expression of circRNAs in the peripheral blood of 6 patients with AS and 6 healthy controls (HC). To screen the differentially expressed circRNAs by fold change (FC) and P value, these differentially expressed circRNAs were analyzed by bioinformatics. In 60 cases of AS and 30 cases of HC, 4 circRNAs were subjected to real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), and their correlation with various clinical indicators was analyzed. Finally, the receiver operating characteristic (ROC) curve was used to analyze their potential as AS diagnostic markers.@*RESULTS@#The microarray results showed that there were 1369 significantly differently expressed (P  1.5) circRNAs between the AS and HC groups (675 upregulated and 694 downregulated). The results of bioinformatics analysis suggested that they were mainly involved in "enzyme binding," "adenosine ribonucleotide binding," "MAPK signaling pathway", etc. The RT-qPCR results showed that the expressions of hsa_circRNA_001544 (U = 486.5, P < 0.05) and hsa_circRNA_102532 (U = 645, P < 0.05) were significantly different between the AS group and the HC group. The AS group was further divided into two subgroups: active AS (ASA) and stable AS (ASS). After analysis, it was found that compared with the HC group, hsa_circRNA_001544 was significantly increased in both ASA (U = 214, P < 0.05) and ASS groups (U = 273, P < 0.05), while hsa_circRNA_008961 (U = 250, P < 0.05) and hsa_circRNA_102532 (U = 295, P < 0.05) were only significantly increased in the ASA group. Furthermore, hsa_circRNA_012732 was significantly different between the ASA and ASS groups (U = 194, P < 0.05), and there was no statistical significance among the remaining groups. Correlation analysis results showed that hsa_circRNA_012732 was negatively correlated with Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), high-sensitivity C-reactive protein (hsCRP), and globulin (GLOB) and positively correlated with lymphocyte count (LY), mean corpusular volume, and albumin (ALB), and hsa_circRNA_008961 was negatively correlated with platelet (PLT) count. ROC curve analysis showed that hsa_circRNA_001544 (95% CI = 0.610-0.831, P < 0.05) and hsa_circRNA_102532 (95% CI = 0.521-0.762, P < 0.05) were statistically significant, and their area under curve (AUC) values were 0.720 and 0.642, respectively.@*CONCLUSIONS@#There are differentially expressed circRNAs in PBMCs of AS patients, and they may be involved in the occurrence and development of AS. Among these differentially expressed circRNAs, hsa_circRNA_012732 has the potential to become an indicator of disease activity, and hsa_circRNA_001544 has the potential to become a molecular marker for AS diagnosis.


Subject(s)
Humans , Leukocytes, Mononuclear , RNA/genetics , RNA, Circular , ROC Curve , Spondylitis, Ankylosing/genetics
6.
Journal of Central South University(Medical Sciences) ; (12): 1080-1089, 2021.
Article in English | WPRIM | ID: wpr-922587

ABSTRACT

OBJECTIVES@#White matter hyperintensity (WMH) is an important factor leading to cognitive impairment, and the mechanism has not been clarified. In recent years, studies have found that circular RNA (circRNA) has differential expression in cerebrovascular diseases. This study aims to analyze the expression profile of circRNA in peripheral blood mononuclear cell (PBMC) of patients with WMH with cognitive impairment, to screen the differentially expressed circRNA, and to explore the possible role of circRNA in WMH with cognitive impairment.@*METHODS@#CircRNA microarray was used to detect the circRNA expression profile of PBMC in patients with WMH with cognitive impairment, and in patients with WMH without cognitive impairment as well as in normal controls (3 cases each, male to female ratio of 2꞉1). The differentially expressed circRNA in patients with WMH with cognitive impairment was screened. The screening criteria for differentially expressed circRNA was fold change (FC) ≥2.0 (|log@*RESULTS@#Compared with the control group, there were 5 significantly up-regulated circRNA and 3 down-regulated circRNA in the WMH with cognitive impairment group; 8 circRNA were significantly up-regulated and 2 were down-regulated in the WMH without cognitive impairment group. When compared with the WMH with cognitive impairment group, no co-differentially expressed circRNA was found in WMH without cognitive impairment group and control group. Compared with the control group, the expression of hsa_circ_0092222 was up-regulated and the expressions of hsa_circ_0000662 and hsa_circ_0083773 were down-regulated in the WMH with cognitive impairment group and the WMH without cognitive impairment group, and there was no significant difference between the 2 groups (all @*CONCLUSIONS@#The circRNA expression profile of patients with WMH is changed significantly. The differentially expressed circRNA may be the cause of WMH; Hsa_circ_0092222, hsa_circ_0000662, and hsa_circ_0083773 may regulate the expression of target genes by targeting adsorption of the target miRNA, leading to brain white matter damage through Janus kinase 2 (JAK2)/signal transducers and activators of transcription (STAT3) signal pathway and Wnt signal pathway.There is no significant difference in circRNA expression profile between WMH with or without cognitive impairment. Cognitive impairment in patients with WMH may have other reasons.


Subject(s)
Female , Humans , Male , Cognitive Dysfunction/genetics , Leukocytes, Mononuclear , MicroRNAs , RNA/genetics , RNA, Circular , Software , White Matter
7.
Braz. j. med. biol. res ; 51(12): e7811, 2018. tab, graf
Article in English | LILACS | ID: biblio-974254

ABSTRACT

Among the novel class of endogenous long non-coding RNAs, circular RNA (circRNA) is known as a key regulator in the development and progression of different cancers. Its function and mechanism in the tumorigenesis of colorectal cancer, however, has not been well studied. This study thus aimed to investigate potential regulation of colorectal cancer by circRNAs and the corresponding regulatory mechanism. We demonstrated that the expression of circRNA hsa_circ_0000523 (also known as circ_006229) was down-regulated in different colorectal cancer cell lines. It was also found that interference of hsa_circ_0000523 induced proliferation and suppressed apoptosis of colorectal cancer cells, the proliferation rate of which was reduced by the overexpression of hsa_circ_0000523. In addition, we found that miR-31 could recognize hsa_circ_0000523 sequence and that it acted as a "sponge" of miR-31, indirectly regulating Wnt/β-catenin signaling pathway, which was involved in the progression of colorectal cancer. The results suggested that the expression of hsa_circ_0000523 correlated to the tumorigenesis of colorectal cancer cells. In addition, as a sponge of miR-31, the low level of hsa_circ_0000523 led to activation of Wnt/β-catenin signaling pathway, inducing the subsequent progress of colorectal cancer.


Subject(s)
Humans , RNA/physiology , Colorectal Neoplasms/genetics , Gene Expression Regulation, Neoplastic/genetics , Apoptosis/genetics , MicroRNAs/genetics , Cell Proliferation/genetics , RNA/genetics , RNA, Neoplasm/genetics , Cell Line, Tumor , RNA, Circular
8.
Mem. Inst. Oswaldo Cruz ; 112(8): 572-576, Aug. 2017. graf
Article in English | LILACS | ID: biblio-1040575

ABSTRACT

The Telomeric Repeat-containing RNAs (TERRA) participate in the homeostasis of telomeres in higher eukaryotes. Here, we investigated the expression of TERRA in Leishmania spp. and Trypanosoma brucei and found evidences for its expression as a specific RNA class. The trypanosomatid TERRA are heterogeneous in size and partially polyadenylated. The levels of TERRA transcripts appear to be modulated through the life cycle in both trypanosomatids investigated, suggesting that TERRA play a stage-specific role in the life cycle of these early-branching eukaryotes.


Subject(s)
Trypanosoma brucei brucei/genetics , RNA/genetics , Repetitive Sequences, Nucleic Acid/genetics , Telomerase/genetics , Leishmania/genetics
9.
Journal of Veterinary Science ; : 163-170, 2012.
Article in English | WPRIM | ID: wpr-174788

ABSTRACT

The purpose of this study was to measure the thickness of canine epidermis at various anatomical sites according to localization of cornified envelopes (involucrin and filaggrin), keratins (keratin 10, 5), and their mRNA expression. This was done in the skin of five breeds of dogs including seven poodles, six golden retrievers, six Shih Tzus, four pugs, and four Labrador retrievers. Epidermal thickness of the stratum corneum and nucleated epidermal layer was significantly different. The greatest thickness was observed in the digital web area and the thinnest epidermis was in the axilla. Epidermal thickness was also significantly different between the breeds (p < 0.05). Immunohistochemical staining scores revealed significant decreases of involucrin, filaggrin, and keratin 10 in the ventral and weight-bearing sites, and a relative increase of keratin 5 (p < 0.05). q-PCR analysis showed that their the levels of mRNA were positively correlated with expression of the corresponding proteins in skin samples (p < 0.05). The present study is the first to report the relationship between epidermal gene expression and histologic morphology of the skin in normal dogs. Further studies will be essential to fully understand the pathogenesis of skin barrier dysfunctions in canines.


Subject(s)
Animals , DNA, Complementary/genetics , Dogs/anatomy & histology , Gene Expression Regulation/physiology , Intermediate Filament Proteins/genetics , Keratin-10/genetics , Keratin-5/genetics , Polymerase Chain Reaction/methods , Protein Precursors/genetics , RNA/genetics , Skin/anatomy & histology
10.
Journal of Forensic Medicine ; (6): 455-459, 2011.
Article in Chinese | WPRIM | ID: wpr-983701

ABSTRACT

With the development of molecular biology, the evidences of genetics has been used widely in forensic sciences. DNA technology has played an important role in individual identification and paternity testing, RNA technology is showing more and more wide application in prospect. This article reviews the application and progress of RNA in forensic science including estimation of postmortem interval, bloodstain age, wound age, as well as determination of cause of death and the source of body fluids.


Subject(s)
Animals , Humans , Actins/metabolism , Blood Stains , Body Fluids/metabolism , Cause of Death , Forensic Medicine/methods , Gene Expression , Genetic Markers/genetics , MicroRNAs/genetics , Polymerase Chain Reaction/methods , Postmortem Changes , RNA/genetics , RNA Stability , RNA, Messenger/metabolism , RNA, Ribosomal, 18S/genetics
11.
Experimental & Molecular Medicine ; : 30-37, 2010.
Article in English | WPRIM | ID: wpr-104281

ABSTRACT

The cytoplasmic polyadenylation element (CPE)-binding protein (CPEB) binds to CPE containing mRNAs on their 3' untranslated regions (3'UTRs). This RNA binding protein comes out many important tasks, especially in learning and memory, by modifying the translational efficiency of target mRNAs via poly (A) tailing. Overexpressed CPEB has been reported to induce the formation of stress granules (SGs), a sort of RNA granule in mammalian cell lines. RNA granule is considered to be a potentially important factor in learning and memory. However, there is no study about RNA granule in Aplysia. To examine whether an Aplysia CPEB, ApCPEB1, forms RNA granules, we overexpressed ApCPEB1-EGFP in Aplysia sensory neurons. Consistent with the localization of mammalian CPEB, overexpressed ApCPEB1 formed granular structures, and was colocalized with RNAs and another RNA binding protein, ApCPEB, showing that ApCPEB1 positive granules are RNA-protein complexes. In addition, ApCPEB1 has a high turnover rate in RNA granules which were mobile structures. Thus, our results indicate that overexpressed ApCPEB1 is incorporated into RNA granule which is a dynamic structure in Aplysia sensory neuron. We propose that ApCPEB1 granule might modulate translation, as other RNA granules do, and furthermore, influence memory.


Subject(s)
Animals , Aplysia/genetics , Fluorescence Recovery After Photobleaching , RNA/genetics , Sensory Receptor Cells/metabolism , mRNA Cleavage and Polyadenylation Factors/genetics
12.
Korean Journal of Ophthalmology ; : 244-250, 2007.
Article in English | WPRIM | ID: wpr-171841

ABSTRACT

PURPOSE: Hepatocyte growth factor (HGF) and its receptor (HGFR/c-Met) regulate motility, mitogenesis, and morphogenesis in a cell type-dependent fashion. We report the role of HGF and c-Met on stress-induced ARPE-19 human retinal pigment epithelial (RPE) cells in this study. METHODS: The cells were cultured either with or without serum. Southern and Western blot analyses were done to determine the expression patterns of HGF/c-Met in serum-starved ARPE-19 cells. The cell proliferation pattern in serum-starved condition was analyzed using MTS assay. Inhibition level of cell proliferation was analyzed using a neutralizing monoclonal antibody against c-Met (2 microgram/ml). RESULTS: Abnormal cell proliferation and scattering of ARPE-19 cells was observed under serum starvation. HGF/c-Met were expressed in serum-starved ARPE-19 cells. ARPE-19 cell proliferation was also enhanced with recombinant HGF treatment. Neutralization against c-Met inhibited the proliferation of serum-deprived ARPE-19 by 64.5% (n=9, S.D. 5.5%). Serum starvation appears to induce epithelial-mesenchymal transition of ARPE-19 cells, resulting in scatter, and the expression of alpha-smooth muscle actin (alpha-SMA), a marker for fibrosis. CONCLUSIONS: In conclusion, c-Met induced under non-physiologic conditions has significant effects on the activation of RPE cells.


Subject(s)
Humans , Blotting, Southern , Blotting, Western , Cell Movement/physiology , Cell Proliferation , Cells, Cultured , Culture Media, Serum-Free , Gene Expression , Hepatocyte Growth Factor/biosynthesis , Mitosis/physiology , Pigment Epithelium of Eye/cytology , Polymerase Chain Reaction , Proto-Oncogene Proteins c-met/biosynthesis , RNA/genetics
13.
J Genet ; 2006 Dec; 85(3): 171-8
Article in English | IMSEAR | ID: sea-114491

ABSTRACT

Some members of hairy/Enhancer-of-split-related gene (HES) family have important effects on axial mesoderm segmentation and the establishment and maintenance of the somite fringe. In fishes, the her6 gene, a member of the HES family, is the homologue of hes1 in mammals and chicken. In this study, the her6 gene and its full-length cDNA from the common carp (Cyprinus carpio) were isolated and characterized. The genomic sequence of common carp her6 is approximately 1.7 kb, with four exons and three introns, and the full-length cDNA of 1314 bp encodes a putative polypeptide of 271 amino acids. To analyse the promoter sequence of common carp her6, sequences of various lengths upstream from the transcription initiation site of her6 were fused to enhanced green fluorescent protein gene (eGFP) and introduced into zebrafish embryos by microinjection to generate transgenic embryos. Our results show that the upstream sequence of 500 bp can direct highly efficient and tissue-specific expression of eGFP in zebrafish embryos, whereas a fragment of 200 bp containing the TATA box and a partial suppressor of hairless paired site sequence (SPS) is not sufficient to drive eGFP expression in zebrafish embryos.


Subject(s)
3' Untranslated Regions , 5' Untranslated Regions , Amino Acid Sequence , Animals , Animals, Genetically Modified , Base Sequence , Chromosome Mapping , Chromosomes , Cloning, Molecular/methods , DNA/genetics , DNA, Complementary/genetics , Embryo, Nonmammalian , Enhancer Elements, Genetic , Exons , Genes, Reporter , Genome , Green Fluorescent Proteins/metabolism , Introns , Microinjections , Molecular Sequence Data , Oocytes/cytology , Polymerase Chain Reaction , Promoter Regions, Genetic , Protein Structure, Tertiary , RNA/genetics , Recombinant Fusion Proteins/chemistry , Regulatory Sequences, Nucleic Acid , Sequence Homology, Amino Acid , Transcription Initiation Site
14.
Journal of Huazhong University of Science and Technology (Medical Sciences) ; (6): 741-3, 2005.
Article in English | WPRIM | ID: wpr-634335

ABSTRACT

To detect the expression of telomerase subunits (human telomerase reverse transcriptase, human telomerase associated protein 1 and human telomerase RNA) in gastric cancer and to examine the role that different telomerase subunits play in the gastric carcinogenesis, reverse transcription-polymerase chain reaction (RT-PCR) was used to detect telomerase subunits messenger RNA in 24 samples of gastric cancer and corresponding non-cancerous tissue. The results showed that the positive rate of hTERT mRNA from gastric cancer and corresponding non-cancerous tissues was 100% and 25%, respectively. The former was significantly higher than the latter (chi2 = 26.4, P 0.05). The positive rates of hTR for gastric cancer and corresponding non-cancerous tissues were both 100% and no significant difference existed between them. It is concluded that in contrast to hTEP1 and hTR, the up-regulation of hTERT mRNA expression may play a more important role in the development of gastric cancer.


Subject(s)
Carrier Proteins/biosynthesis , Carrier Proteins/genetics , RNA/biosynthesis , RNA/genetics , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Reverse Transcriptase Polymerase Chain Reaction , Stomach Neoplasms/metabolism , Telomerase/biosynthesis , Telomerase/genetics
15.
J Biosci ; 2002 Mar; 27(2): 165-86
Article in English | IMSEAR | ID: sea-110855

ABSTRACT

We have investigated the origin of genes, the genetic code, proteins and life using six indices (hydropathy, alpha-helix, beta-sheet and beta-turn formabilities, acidic amino acid content and basic amino acid content) necessary for appropriate three-dimensional structure formation of globular proteins. From the analysis of microbial genes, we have concluded that newly-born genes are products of nonstop frames (NSF) on antisense strands of microbial GC-rich genes [GC-NSF(a)] and from SNS repeating sequences [(SNS)n] similar to the GC-NSF(a) (S and N mean G or C and either of four bases, respectively). We have also proposed that the universal genetic code used by most organisms on the earth presently could be derived from a GNC-SNS primitive genetic code. We have further presented the [GADV]-protein world hypothesis of the origin of life as well as a hypothesis of protein production, suggesting that proteins were originally produced by random peptide formation of amino acids restricted in specific amino acid compositions termed as GNC-, SNS- and GC-NSF(a)-0th order structures of proteins. The [GADV]-protein world hypothesis is primarily derived from the GNC-primitive genetic code hypothesis. It is also expected that basic properties of extant genes and proteins could be revealed by considerations based on the scenario with four stages.


Subject(s)
Amino Acids/metabolism , Animals , Biological Evolution , Evolution, Molecular , Exons/genetics , Genes , Genetic Code , Mitochondria/genetics , Models, Genetic , Protein Structure, Secondary , Proteins/chemistry , RNA/genetics
16.
Southeast Asian J Trop Med Public Health ; 2002 ; 33 Suppl 3(): 139-44
Article in English | IMSEAR | ID: sea-35815

ABSTRACT

We developed in-house RNA extraction and RT-PCR reagent kits for the molecular serotyping of dengue viruses in field-caught Aedes mosquitos. Mosquitos that showed positive results by ELISA or IFA were selected for the identification of dengue viruses in order to predict the distribution of the four dengue serotypes. Total RNA was extracted from one whole mosquito as well as from one dissected mosquito by our in-house RNA extraction reagents using the modified method of guanidinium thiocyanate denaturation and isopropanol precipitation. The extracted RNA was amplified by our in-house RT-PCR reagents specific for each dengue serotype under optimized conditions. Dengue viral RNA extracted from a single mosquito as well as from the head and thorax of one dissected mosquito could be detected successfully; it could not be found in the abdomen, however. These results indicated that most of the dengue viruses were located in the head and thorax rather than in the abdomen. The results of dengue serotyping showed a pure specific PCR product for each dengue serotype at 490, 230, 320 and 398 bp for DEN-1, DEN-2, DEN-3, and DEN-4 respectively. In addition, the detection sensitivity was very high: an amount of RNA template equivalent to approximately 1/80 of a single mosquito could be detected by agarose gel electrophoresis and ethidium bromide staining. The coupling of RT-PCR-based surveillance of dengue viral infection with disease mapping data (Geograpical Information System, GIS) could serve as an alternative epidemiological means of providing an early warning of dengue fever/dengue hemorrhagic fever epidemics.


Subject(s)
Animals , Dengue Virus/genetics , Housing , Indicators and Reagents , RNA/genetics , Reverse Transcriptase Polymerase Chain Reaction , Sensitivity and Specificity
17.
J Biosci ; 2001 Mar; 26(1): 25-38
Article in English | IMSEAR | ID: sea-111291

ABSTRACT

The nucleus-limited large non-coding hsr(omega)-n RNA product of the 93D or the hsr(omega) gene of Drosophila melanogaster binds to a variety of RNA-binding proteins involved in nuclear RNA processing. We examined the developmental and heat shock induced expression of this gene by in situ hybridization of nonradioactively labelled riboprobe to cellular transcripts in intact embryos, larval and adult somatic tissues of wild type and an enhancer-trap line carrying the hsr(omega) 05241 allele due to insertion of a P-LacZ-rosy+ transposon at -130 bp position of the hsr(omega) promoter. We also examined LacZ expression in the enhancer-trap line and in two transgenic lines carrying different lengths of the hsr(omega) promoter upstream of the LacZ reporter. The hsr(omega) gene is expressed widely at all developmental stages; in later embryonic stages, its expression in the developing central nervous system was prominent. In spite of insertion of a big transposon in the promoter, expression of the hsr(omega) 05241 allele in the enhancer-trap line, as revealed by in situ hybridization to hsr(omega) transcripts in cells, was similar to that of the wild type allele in all the embryonic, larval and adult somatic tissues examined. Expression of the LacZ gene in this enhancer-trap line was similar to that of the hsr(omega) RNA in all diploid cell types in embryos and larvae but in the polytene cells, the LacZ gene did not express at all, neither during normal development nor after heat shock. Comparison of the expression patterns of hsr(omega) gene and those of the LacZ reporter gene under its various promoter regions in the enhancer-trap and transgenic lines revealed a complex pattern of regulation, which seems to be essential for its dynamically varying expression in diverse cell types.


Subject(s)
Animals , Drosophila melanogaster/genetics , Gene Expression Regulation, Developmental , Genes, Reporter , Heat-Shock Proteins/genetics , In Situ Hybridization , Lac Operon , Larva/metabolism , Promoter Regions, Genetic , RNA/genetics
18.
J Biosci ; 2000 Dec; 25(4): 361-6
Article in English | IMSEAR | ID: sea-111018

ABSTRACT

Mosquitoes are vectors for the transmission of many human pathogens that include viruses, nematodes and protozoa. For the understanding of their vectorial capacity, identification of disease carrying and refractory strains is essential. Recently, molecular taxonomic techniques have been utilized for this purpose. Sequence analysis of the mitochondrial 16S rRNA gene has been used for molecular taxonomy in many insects. In this paper, we have analysed a 450 bp hypervariable region of the mitochondrial 16S rRNA gene in three major genera of mosquitoes, Aedes, Anopheles and Culex. The sequence was found to be unusually A+T rich and in substitutions the rate of transversions was higher than the transition rate. A phylogenetic tree was constructed with these sequences. An interesting feature of the sequences was a stretch of Ts that distinguished between Ae-des and Culex on the one hand, and Anopheles on the other. This is the first report of mitochondrial rRNA sequences from these medically important genera of mosquitoes.


Subject(s)
Animals , Base Sequence , Culicidae/genetics , Evolution, Molecular , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , Polymorphism, Genetic , Polymorphism, Restriction Fragment Length , RNA/genetics , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Species Specificity
19.
Braz. j. infect. dis ; 3(4): 156-62, Aug. 1999. ilus, tab
Article in English | LILACS | ID: lil-254771

ABSTRACT

During April, 1997, to February, 1998, an epidemiological survey of group A rotavirus strains causing acute diarrhea in infants and young children was conducted in Hospital Municipal Jesus (Rio de Janeiro, D.C.). Rotaviruses were detected in 18.6 percent of examined specimens. Among the rotavirus strains tested for RNA by PAGE, 70 percent belonged to the long pattern and 30 percent the short pattern. A long pattern classified as LD was the most common electropherotype among all of the types. This same pattern had been identified in the city 15 years ago. The most common short pattern was SA type and a few were SB types. The SB types were more common than SA types 15 years ago. Rotavirus infection was observed throughout the period studied, but peaks of infection occurred in May, June, and September. The long pattern was recorded continuously and in all ages studied (0-8 years old), but the short pattern ocurred only in young children (age range 0-11 months) and only in May and June, 1997. This data show epidemiologic differences in infection associated with long and short electropherotypes; the long electropherotypes appear to circulate continuously in the community, whereas the sort electropherotype strains appear only in young children in an episodic fashion.


Subject(s)
Humans , Infant, Newborn , Infant , Child, Preschool , Child , Diarrhea, Infantile/diagnosis , Diarrhea, Infantile/epidemiology , Electrophoresis, Polyacrylamide Gel , Rotavirus Infections/epidemiology , Rotavirus/genetics , Rotavirus/isolation & purification , Brazil/epidemiology , Enzyme-Linked Immunosorbent Assay , Molecular Epidemiology , RNA/genetics
SELECTION OF CITATIONS
SEARCH DETAIL