Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The aim of this study was the investigation of hepatitis C virus (HCV) RNA by reverse transcriptase-polymerase chain reaction (RT-PCR), in hemodialysis patients in order to detect HCV infection, and the determination of virus genotypes in HCV positive patients. A total of 94 patients, 44 female and 50 male (mean age: 49 +/- 17 years old), who were included in hemodialysis program between 1987-1999 (0-13 years) in the Hemodialysis Units of Gazi University, School of Medicine and Ankara Etlik Hospital of Social Insurance Society, were enrolled in the study. Anti-HCV was found positive in 41 patients (43.6%), while HCV-RNA was positive in 36 patients (38.3%). Thirty three of the patients were positive for both anti-HCV and HCV-RNA, 8 were anti-HCV positive HCV-RNA negative, and 3 were anti-HCV negative HCV-RNA positive. HCV was genotyped by Restriction Fragment Length Polymorphism (RFLP) method in 36 HCV-RNA positive patients, and 28 yielded genotype 1b, and 8 showed genotype 1a pattern. PCR products from 5'UTR of HCV-RNA isolated from two patients were also sequenced to confirm the results of RFLP study. The rate of anti-HCV and HCV-RNA positivity was detected statistically higher in patients who exposed hemodialysis longer than 24 months, and in hemodialysis patients who had blood transfusion than the other groups. As a result, our findings supported that the periodic follow-up of HCV-RNA, in addition to anti-HCV would be useful for hemodialysis patients, and genotyping of HCV for the epidemiologic data.
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PMID:[The presence of hepatitis C virus (HCV) infection in hemodialysis patients and determination of HCV genotype distribution]. 1283 63

Gonadotropin-releasing hormone (GnRH), which is essential for reproductive function, is made by neurones that migrate from the nasal region into the brain during early embryonic development. This migration begins in chick when the olfactory pit is formed. This is approximately the time that GnRH neurones can be detected immunocytochemically. The present study investigated (i). how early in development the GnRH gene is expressed and (ii). the sites of its expression. Accordingly, reverse transcriptase-polymerase chain reaction (PCR) and in situ hybridization were performed on chick embryos before gastrulation up until the stage by which GnRH neurones have begun to migrate into the central nervous system. Primers were made to the 5'- and 3'-UTR region of the message for cGnRH-I, the form of the peptide that is essential for reproductive function in the chicken. PCR product was found in all stages and the sequences of products from all stages were identical. Thus, the GnRH gene is expressed continuously throughout embryonic development. In situ hybridization with a digoxygenin labelled riboprobe revealed staining along the primitive streak immediately before gastrulation. In later stages, cGnRH-I gene expression was seen in association with the anterior neural ridge. The expression was subsequently restricted to a narrow, clearly defined region, which is associated with the presumptive nasal cavity and olfactory placode. Later, GnRH neurones could be seen in their migratory routes by both in situ hybridization and immunocytochemistry. Expression of the GnRH gene has been described in preimplantation stages in mammals and there is evidence that the neuropeptide plays a role in formation and maintenance of the placenta. What role (if any) it may play in early avian development remains unknown. The demonstration of sites of GnRH expression during the early period of neurulation suggests that GnRH neurones arise before olfactory placode formation.
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PMID:Early expression of chicken gonadotropin-releasing hormone-1 in the developing chick. 1289 81

GB virus C (GBV-C) is a virus that has been proposed as a member of the Flaviviridae family, distantly related to hepatitis C virus (HCV). The virus is able to infect humans parenterally and perinatally, although its true pathogenicity remains unknown. The 5' terminal region of GBV-C is the most highly conserved region of the virus genome. Comparison of 5' untranslated region (5' UTR) sequences from GBV-C infected individuals shows that variation is limited to particular sites that are often covariant and associated with different virus genotypes. Extensive sequence analysis of the GBV-C genome provides evidence for the existence of at least five major genotypes, some of which can be further divided into subtypes. For genotyping by restriction fragment length polymorphism (RFLP), it is essential to identify genomic positions that not only reflect genotype differences, but that also harbor restriction sites that allow recognition of these differences. Restriction site analysis of type-specific sequence motifs predicted that endonucleases BsmFI, HaeII, HinfI, and ScrFI could be used for the identification all known genotypes (types 1-5) with 99.6% accuracy. The method was applied to serum samples from 46 chronic GBV-C carriers of heterogeneous geographical and ethnic origin, comparing observed cleavage patterns of GBV-C variants amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) of the 5' UTR with the RFLP predicted from sequences deposited in GenBank database. cDNA sequencing and subsequent alignment of the 46 GBV-C isolates confirmed RFLP profiles predicted theoretically. The observed geographical distribution of genotypes is also in agreement with previous reports. This method may be useful for rapid and reliable characterization of GBV-C isolates when either epidemiological or transmission studies are carried out.
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PMID:Genotyping of GB virus C by restriction pattern analysis of the 5' untranslated region. 1293 97

The expression of cytochrome P450scc, encoded by the CYP11A gene, was investigated in the rat kidney from birth to adulthood. In the male and female rat kidneys, the corresponding mRNA was detected by semi-quantitative reverse transcriptase and polymerase chain reaction (RT-PCR) analysis with specific primers, resulting in higher levels of expression during the first 15 days from birth. RT-PCR and sequence analysis showed that the P450scc mRNA coding region was the same for both kidney and testis, whereas 5'-RACE analysis (rapid amplification of cDNA ends) demonstrated that the renal transcription utilizes a distal transcription start site (TSS) located 76 b upstream of that used in ovarian and testicular P450scc mRNA expression, which is placed 43 b upstream of the first ATG. The 5'-UTR sequence of renal P450scc cDNA exactly matched the contiguous upstream untranslated region of the gene, suggesting that alternative splicing was not involved in the synthesis of this transcript. Northern hybridization detected a specific transcript only in the newborn male, but not in adult rat kidney, confirming the higher levels of expression in the first days of the rat's life. Positive immunodetections of cytochrome P450scc were found in renal cortical distal tubules and the results were confirmed by Western blotting analysis. As demonstrated by semi-quantitative RT-PCR, the male kidney also expresses the messengers corresponding to the steroidogenic acute regulatory (StAR) and steroidogenic factor 1 (SF-1) proteins, which are normally required for steroidogenesis in steroidogenic tissues, such as gonads and adrenal cortex. These studies suggest that the rat kidney has the capability for local steroid hormone production, although the physiological significance of the pregnenolone eventually produced remains to be established.
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PMID:Expression of cytochrome P450scc mRNA and protein in the rat kidney from birth to adulthood. 1502 86

The development of single, sensitive, fluorogenic reverse transcriptase-polymerase chain reaction (TaqMan) assays were required for the rapid and specific detection of three encephalitic viruses found in the Australasian region, namely; Japanese encephalitis virus (JEV), Murray Valley encephalitis virus (MVEV), and Kunjin virus (KUNV). Primers and a fluorogenic probe were individually designed to be complementary to a nucleotide region encompassing the 3' terminus of the nonstructural (NS) 5 gene and a portion of the 3' untranslated region (NS5-3'UTR) of each of the viral genomes respectively. Synthetically produced primer and probe controls were developed to minimize the likelihood of contamination and generation of false positives. Viral RNA from singly infected mosquitoes could be detected in pools of 1000 mosquitoes and positive mosquito pools collected from the field have been identified using each assay, indicating a high level of sensitivity and suitability for use in mosquito surveillance programs. In addition, the JEV TaqMan assay has been used to detect successfully viral RNA in sentinel pig serum samples. These assays potentially offer superior and timely detection of encephalitic viruses from surveillance samples, which is essential for the rapid implementation of vector control measures and continued monitoring of virus activity in the Australasian region.
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PMID:Detection of Australasian Flavivirus encephalitic viruses using rapid fluorogenic TaqMan RT-PCR assays. 1504 Dec 13

Glutathione S-transferases (GSTs) are scavenging enzymes that detoxify cellular xenobiotics and toxins by catalyzing the conjugation of these substrates with a tripeptide glutathione. GSTs are classified depending on gene organization and sequence similarity. The sequence analysis of genomic DNA for zeta class GST (GSTZ) locus in rice indicated that two homologous GSTZ genes lay in a tandem orientation with a short (0.4 kb) intergenic spacer. The upstream OsGSTZ1 and downstream OsGSTZ2 spanned 3.5 and 3.2 kb with nine coding exons, respectively. The transcript of OsGSTZ1 had a long 3' untranslated region (3' UTR) that was mostly encoded by a 10th noncoding exon, whereas OsGSTZ2 mRNA contained a long 5' UTR. Northern blot analysis showed that OsGSTZ1/2 messages were strongly expressed in leaf blades, while transcripts from roots were low level. Because OsGSTZ1/2 messages in leaf tissues were strongly induced only by water treatment, it was difficult to assay for the induction of OsGSTZ1/2 transcripts by various stress treatments. Thus, using rice culture cells, we analyzed the respective responses of OsGSTZ1 and OsGSTZ2 genes against various treatments by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). The results showed that OsGSTZ1 was expressed at a level ca. 1000-fold higher than OsGSTZ2 in suspension cells without stress treatment. OsGSTZ1 was expressed constitutively under various stress conditions. In contrast, the expression of OsGSTZ2 gene was strongly enhanced to 30-fold by treatment with jasmonic acid. These observations suggested that the expression of OsGSTZ1 and OsGSTZ2 genes are differentially regulated in the culture cell of rice.
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PMID:Genomic structure and differential expression of two tandem-arranged GSTZ genes in rice. 1519 97

An investigation was initiated to explore previously published results indicating that approximately 80 bp of the 5'-end of the iduronate sulfatase (IDS) cDNA sequence (Accession No L07291) are 100% homologous with the 3'-UTR of isoform I of the sodium hydrogen exchanger (Acc. No. U51112). 5'-RACE carried out on IDS mRNA demonstrated the apparent homology to be a cloning artifact. A sequence comparison of the IDS 5'-RACE product with a mouse BAC clone covering the region, and with various IDS ESTs, suggested that the region is highly susceptible to cloning artifacts, a common one of which is template switching by reverse transcriptase. The nucleotide sequence flanking the translation start site is unusual in containing two inverted repeats composed of the complementary trinucleotide microsatellites, (GCG)9 and (CGC)6. These likely form a highly stable stem of 20-21 nt, through which reverse transcription is compromised. Such a stem could be involved in the regulation of IDS expression by directly affecting translation, message turnover, or serving as a substrate for siRNA production. Though such mRNA features are relatively rare, they may be more abundant but overlooked due to difficulties in their reverse transcription.
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PMID:Inverted Gcg/CGC trinucleotide microsatellites in the 5'-region of Mus IDS mRNA: recurrent induction of aberrant reverse transcripts. 1529 86

This work describes 18 new transcribed retrotransposons of the blood fluke Schistosoma mansoni. Among them, 9 were LTR, 8 non-LTR, and 1 Penelope-like element (PLE) retrotransposon. Sequences were generated by in silico reconstruction using S. mansoni ESTs and transcripts obtained by rapid amplification of cDNA ends, complemented in some cases by sequencing of genomic clones amplified by PCR. A novel element from the ancient R2/R4/CRE transposon group is described for the first time in S. mansoni. In addition, one non-LTR retrotransposon family displays long (40-450 bp) 3'-UTR with at least six different transcribed sequences among the copies, five LTR retrotransposons have abundantly transcribed incomplete copies lacking the sequence segment coding for the reverse transcriptase domain, and four non-LTR retrotransposons code for DNA-binding PHD domains that may give them a differential targeting. These results allow for a comprehensive description of the transcribed retrotransposon diversity of this complex human parasite.
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PMID:Identification of 18 new transcribed retrotransposons in Schistosoma mansoni. 1593 96

The objective of this study was to identify single nucleotide polymorphisms (SNPs) within four functionally related immune response genes in the horse, and to develop genotyping techniques that could be useful for future genomic studies of horse infectious and allergic diseases. The genes analysed were: the lipopolysaccharide (LPS) receptor gene CD14, the toll-like receptor 4 gene TLR4, the gene Cepsilon encoding the IgE heavy chain molecule and the gene FcepsilonR1 alpha coding for the alpha subunit of the IgE receptor molecule. Horse-specific primers amplifying selected gene regions were designed and SNPs were searched by selective resequencing and/or by PCR-SSCP (polymerase chain reaction-sequence specific conformational polymorphism) or PCR-RFLP (PCR-restriction fragment length polymorphism). Gene expression was analysed by RT-PCR (reverse transcriptase-PCR) of all four genes examined. For CD14, the cDNA sequence was determined and a novel sequence of the 5'UTR region was identified. The protein-coding sequence was identical to that previously deposited in GenBank. 5'UTR, intronic and both synonymous and non-synonymous exonic SNPs were identified. Three SNPs were found in the CD14 gene, four in the TLR4 gene; two SNPs were identified in the Cepsilon gene, and one SNP was found in the FcepsilonR1 alpha gene. PCR-RFLP was developed for genotyping eight of the SNPs identified. The RT-PCR assay showed that all the SNPs reported here are parts of expressed genes. The results showed that important immunity-related genes in horses are polymorphic and that even non-synonymous SNPs with potential functional impact may occur. The methods developed for genotyping and haplotyping the SNPs identified represent, along with markers described previously, a potentially useful tool for genomic analysis of the function and role of these genes in immunity and in mechanisms of disease.
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PMID:Single nucleotide polymorphisms in four functionally related immune response genes in the horse: CD14,TLR4, Cepsilon, andFcepsilon R1 alpha. 1616 94

To understand the role of the Tigriopus japonicus Hsp20 gene, we isolated this gene from a whole body cDNA library and found two heat shock factor elements at the 5'-UTR. The transformed bacteria containing Tigriopus Hsp20 showed thermotolerance against heat shock (54 degrees C) with different ranges of time. The Tigriopus Hsp20 gene is comprised of 174 amino acid residues and shows similarity to Caenorhabditis elegans (27% identity), silkworm (24.1% identity), moth (24.1% identity), Mexican tetra (19.5% identity), zebrafish (19.5% identity), and spiny dogfish (17.2% identity) genes, but shows more similarity in the C-terminal region that contains an alpha-crystallin domain. Protein motifs such as an N-glycosylation site (67-70 NKSE) and a casein kinase II phosphorylation site were found in Tigriopus Hsp20. The genomic structure of the Tigriopus Hsp20 gene did not contain introns. To characterize the biochemical characteristics of the Tigriopus Hsp20 protein, we expressed Tigriopus Hsp20 in Escherichia coli and purified the soluble protein via 6x His-tag chromatography. To analyze the gene expression of Tigriopus Hsp20 against environmental stresses (e.g., water temperature and salinity), we performed a semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR). On exposure to different salinities, significant change in the expression of Tigriopus Hsp20 was not observed. However, upon heat shock (30 degrees C), Tigriopus Hsp20 expression was significantly increased, but in the case of cold shock (4 or 10 degrees C), expression was likely downregulated. These findings provide a better understanding of cellular protection mechanisms against environmental stress such as heat shock.
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PMID:The intertidal copepod Tigriopus japonicus small heat shock protein 20 gene (Hsp20) enhances thermotolerance of transformed Escherichia coli. 1640 54


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