Pub Date : 2001-01-01DOI: 10.3109/10425170109024997
Gabriela Silva, C. Rodrigues-Pousada
The nucleotide sequence of a 6940 bp DNA fragment from Desulfovibrio gigas, containing seven ORFs was determined. ORF-1 encodes a probable lipoprotein having high similarities with lytic transglycosylases. ORF-2 encodes a polypeptide that does not show homologies with proteins deposited in the database, but it contains the consensus pattern of class II aminoacyl-tRNA synthetases. The putative protein encoded by ORF-3 possesses high similarities with universal stress response proteins from the UspA family. Northern blot analysis of ORF-3 shows that it is constitutively and abundantly expressed. ORF-4 encodes a probable helix-turn-helix-containing DNA-binding protein, given the presence of the helix-turn-helix motif, characteristic of this class of proteins. Its N-terminal region has high identity with its counterparts from proteins belonging to the RRF2 family. Northern blot analysis shows that ORF-4 is transcribed as a single mRNA in contrast to its orthologue from Desulfovibrio vulgaris. ORF-5 encodes a putative fusion protein as its N- and C-termini show significant homologies with molybdenum formylmethanofuran dehydrogenase and molybdopterin biosynthesis proteins, respectively. ORF-7 encodes a prokaryotic lipoprotein having homologies with multidrug efflux and DNA damage-inducible proteins, and it is constitutively and abundantly expressed.
{"title":"A 6940 bp DNA Fragment from Desulfovibrio gigas Contains Genes Coding for Lipoproteins, Universal Stress Response and Transcriptional Regulator Protein Homologues","authors":"Gabriela Silva, C. Rodrigues-Pousada","doi":"10.3109/10425170109024997","DOIUrl":"https://doi.org/10.3109/10425170109024997","url":null,"abstract":"The nucleotide sequence of a 6940 bp DNA fragment from Desulfovibrio gigas, containing seven ORFs was determined. ORF-1 encodes a probable lipoprotein having high similarities with lytic transglycosylases. ORF-2 encodes a polypeptide that does not show homologies with proteins deposited in the database, but it contains the consensus pattern of class II aminoacyl-tRNA synthetases. The putative protein encoded by ORF-3 possesses high similarities with universal stress response proteins from the UspA family. Northern blot analysis of ORF-3 shows that it is constitutively and abundantly expressed. ORF-4 encodes a probable helix-turn-helix-containing DNA-binding protein, given the presence of the helix-turn-helix motif, characteristic of this class of proteins. Its N-terminal region has high identity with its counterparts from proteins belonging to the RRF2 family. Northern blot analysis shows that ORF-4 is transcribed as a single mRNA in contrast to its orthologue from Desulfovibrio vulgaris. ORF-5 encodes a putative fusion protein as its N- and C-termini show significant homologies with molybdenum formylmethanofuran dehydrogenase and molybdopterin biosynthesis proteins, respectively. ORF-7 encodes a prokaryotic lipoprotein having homologies with multidrug efflux and DNA damage-inducible proteins, and it is constitutively and abundantly expressed.","PeriodicalId":11381,"journal":{"name":"DNA Sequence","volume":"18 1","pages":"229 - 238"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83824918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.3109/10425170109047562
M. Sato, K. Miyado, M. Kimura
M32 [also termed chromatin modifier protein 2 (MOD2)] is a nuclear protein consisting of the condensed chromatin structure (heterochromatin) and considered one of the mammalian homologues of heterochromatin protein 1 (HP1), first isolated as one of the components of heterochromatin in Drosophila. This report presents the isolation and characterization of the 5′-upstream region of the mouse M32 gene containing a promoter region and 5′-untranslated region (5′-UTR) exon. The 5′-upstream region (approximately 0.27 kb starting from the 5′ end of the 5′-UTR exon) of the M32 gene contained neither a TATA box nor a CCAAT box, but possessed potential binding sites for transcription factors such as Spl, H4TF-1, PEA2, PEA3, GSG element and Egr-1, and was highly G/C-rich. The promoter activity of this 5′-upstream region was demonstrated by transfecting its fusion-construct with the E. coli β-galactosidase gene into the F9 mouse teratocarcinoma cell line. The 5′ ends of the mRNA were mapped to at least two positions in the 5′-upstream region. Interestingly, the 5′-upstream region exhibited a high degree of similarity to a portion of heterogeneous nuclear ribonucleo-protein (hnRNP) A2/B1 gene, which is thought to play a role in RNA processing, located in the reverse orientation to the M32 gene, and also to several known ESTs and cDNAs. These findings suggest that the 5′-upstream region of the M32 gene consists of a multiple regulatory complex which probably plays important roles in nuclear function such as chromatin organization and RNA processing.
{"title":"Cloning and Characterization of 5′-Upstream Sequence of the M32 Gene for a Mouse Homologue of Drosophila Heterochromatin Protein 1 (HP1)","authors":"M. Sato, K. Miyado, M. Kimura","doi":"10.3109/10425170109047562","DOIUrl":"https://doi.org/10.3109/10425170109047562","url":null,"abstract":"M32 [also termed chromatin modifier protein 2 (MOD2)] is a nuclear protein consisting of the condensed chromatin structure (heterochromatin) and considered one of the mammalian homologues of heterochromatin protein 1 (HP1), first isolated as one of the components of heterochromatin in Drosophila. This report presents the isolation and characterization of the 5′-upstream region of the mouse M32 gene containing a promoter region and 5′-untranslated region (5′-UTR) exon. The 5′-upstream region (approximately 0.27 kb starting from the 5′ end of the 5′-UTR exon) of the M32 gene contained neither a TATA box nor a CCAAT box, but possessed potential binding sites for transcription factors such as Spl, H4TF-1, PEA2, PEA3, GSG element and Egr-1, and was highly G/C-rich. The promoter activity of this 5′-upstream region was demonstrated by transfecting its fusion-construct with the E. coli β-galactosidase gene into the F9 mouse teratocarcinoma cell line. The 5′ ends of the mRNA were mapped to at least two positions in the 5′-upstream region. Interestingly, the 5′-upstream region exhibited a high degree of similarity to a portion of heterogeneous nuclear ribonucleo-protein (hnRNP) A2/B1 gene, which is thought to play a role in RNA processing, located in the reverse orientation to the M32 gene, and also to several known ESTs and cDNAs. These findings suggest that the 5′-upstream region of the M32 gene consists of a multiple regulatory complex which probably plays important roles in nuclear function such as chromatin organization and RNA processing.","PeriodicalId":11381,"journal":{"name":"DNA Sequence","volume":"363 1","pages":"106 - 97"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82990016","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.3109/10425170109084473
Junlin Wu, Anwara . Khan, C. Shih, D. S. Shih
Osmotin and osmotin-like proteins (OLPs) are pathogenesis-related (PR) proteins, whose synthesis is normally stimulated upon infection of plants by pathogens. A strawberry genomic clone containing an osmotin-like protein (OLP) gene was isolated and sequenced. This clone contains an open reading frame of 681 nucleotides without any intron. The predicted amino acid sequence of the protein shares high degrees of homology with a number of other OLPs and related proteins, of which several are known to have antifungal activities. Southern hybridization analysis of strawberry genomic DNA suggested that the OLP is coded by a multi-gene family. Results from reverse transcriptase-polymerase chain reaction indicated that this OLP gene is expressed in uninfected strawberry plants.
{"title":"Cloning and Sequence Determination of a gene Encoding an Osmotin-like Protein from Strawberry (Fmgaria X ananassa Duch.)","authors":"Junlin Wu, Anwara . Khan, C. Shih, D. S. Shih","doi":"10.3109/10425170109084473","DOIUrl":"https://doi.org/10.3109/10425170109084473","url":null,"abstract":"Osmotin and osmotin-like proteins (OLPs) are pathogenesis-related (PR) proteins, whose synthesis is normally stimulated upon infection of plants by pathogens. A strawberry genomic clone containing an osmotin-like protein (OLP) gene was isolated and sequenced. This clone contains an open reading frame of 681 nucleotides without any intron. The predicted amino acid sequence of the protein shares high degrees of homology with a number of other OLPs and related proteins, of which several are known to have antifungal activities. Southern hybridization analysis of strawberry genomic DNA suggested that the OLP is coded by a multi-gene family. Results from reverse transcriptase-polymerase chain reaction indicated that this OLP gene is expressed in uninfected strawberry plants.","PeriodicalId":11381,"journal":{"name":"DNA Sequence","volume":"77 12","pages":"447 - 453"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91547077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.3109/10425170109080772
Yann Gibert, S. Spada, J. Wall, J. Pembroke
Screening of a genomic library of the thermophile Thermus thermophilus revealed a novel thermophilic hint gene, homologues of which are highly conserved in genera from archaea to mammals. Hint belongs to the HIT protein super-family, which contains two broad groups, Fhit, associated with tumour suppression in eukaryotes and Hint with putatitive protein kinase C inhibitory activity. In T. thermophilus the 321bp gene has a GC content of 67% overall and 94.4% in the third nucleotide position, with unusually no thymine as a wobble base. The gene product, a small highly conserved 11996Da predicted soluble cytoplasmic protein, offers an ideal opportunity to investigate thermostabilising amino acid substitutions. Here we report on the characterisation of the novel hint sequence.
{"title":"Cloning and Characterisation of the Hint Homologue of the Thermophile Thermus thermophilus","authors":"Yann Gibert, S. Spada, J. Wall, J. Pembroke","doi":"10.3109/10425170109080772","DOIUrl":"https://doi.org/10.3109/10425170109080772","url":null,"abstract":"Screening of a genomic library of the thermophile Thermus thermophilus revealed a novel thermophilic hint gene, homologues of which are highly conserved in genera from archaea to mammals. Hint belongs to the HIT protein super-family, which contains two broad groups, Fhit, associated with tumour suppression in eukaryotes and Hint with putatitive protein kinase C inhibitory activity. In T. thermophilus the 321bp gene has a GC content of 67% overall and 94.4% in the third nucleotide position, with unusually no thymine as a wobble base. The gene product, a small highly conserved 11996Da predicted soluble cytoplasmic protein, offers an ideal opportunity to investigate thermostabilising amino acid substitutions. Here we report on the characterisation of the novel hint sequence.","PeriodicalId":11381,"journal":{"name":"DNA Sequence","volume":"26 1","pages":"179 - 185"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81415030","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.3109/10425170109047565
B. Bocquet-Muchembled, R. Leroux, Anne Chotteau-LeliÉGvre, F. Fontaine
Numerous genes belonging to the ets gene family have been described for a few years. The founder of this family is the v-ets proto-oncogene, which is the viral counterpart of the chicken c-ets-1 proto-oncogene. Main research was carried out both on Vertebrates, Drosophila and the nematod Caenorhabditis elegans. Previously, two genes of this family named Nd ets and Nd erg, were isolated in the polychaete annelid Hediste (Nereis) diversicolor. Here we have described the isolation of one gene from the ets family in another polychaete annelid named Perinereis cultrifera. By polymerase chain reaction using degenerated primers, we have amplified an approximatively 515 pb genomic region encoding the ETS domain and another domain designed as “R” domain by Qi et al. (1992) and which can mediate transactivation. By using this method for isolating members of the ets gene family, we are going to realize a phylogenetic study of the phylum of polychaete annelids.
{"title":"Isolation of a Member of ets Gene Family in the Polychaete Annelid Perinereis cultrifera","authors":"B. Bocquet-Muchembled, R. Leroux, Anne Chotteau-LeliÉGvre, F. Fontaine","doi":"10.3109/10425170109047565","DOIUrl":"https://doi.org/10.3109/10425170109047565","url":null,"abstract":"Numerous genes belonging to the ets gene family have been described for a few years. The founder of this family is the v-ets proto-oncogene, which is the viral counterpart of the chicken c-ets-1 proto-oncogene. Main research was carried out both on Vertebrates, Drosophila and the nematod Caenorhabditis elegans. Previously, two genes of this family named Nd ets and Nd erg, were isolated in the polychaete annelid Hediste (Nereis) diversicolor. Here we have described the isolation of one gene from the ets family in another polychaete annelid named Perinereis cultrifera. By polymerase chain reaction using degenerated primers, we have amplified an approximatively 515 pb genomic region encoding the ETS domain and another domain designed as “R” domain by Qi et al. (1992) and which can mediate transactivation. By using this method for isolating members of the ets gene family, we are going to realize a phylogenetic study of the phylum of polychaete annelids.","PeriodicalId":11381,"journal":{"name":"DNA Sequence","volume":"18 1","pages":"121 - 124"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81441201","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.3109/10425170109042052
L. Carim-Todd, L. Sumoy, N. Andreu, Xavier Estivill And, M. Escarceller
We have isolated C15orf5, a novel human gene lacking homology to any known protein. The C15orf5 gene encodes a transcript of 1, 519 nt with an ORF of 94 amino acids and a predicted protein size of 11.5 kDa. Northern blot analysis showed enhanced expression of C15orf5 in heart. C15orf5 was mapped to chromosome 15q23-q24 using the Stanford TNG4 Radiation Hybrid panel.
{"title":"Identification of C15orf5, a Heart-Enriched Transcript on Chromosome 15q23-q24","authors":"L. Carim-Todd, L. Sumoy, N. Andreu, Xavier Estivill And, M. Escarceller","doi":"10.3109/10425170109042052","DOIUrl":"https://doi.org/10.3109/10425170109042052","url":null,"abstract":"We have isolated C15orf5, a novel human gene lacking homology to any known protein. The C15orf5 gene encodes a transcript of 1, 519 nt with an ORF of 94 amino acids and a predicted protein size of 11.5 kDa. Northern blot analysis showed enhanced expression of C15orf5 in heart. C15orf5 was mapped to chromosome 15q23-q24 using the Stanford TNG4 Radiation Hybrid panel.","PeriodicalId":11381,"journal":{"name":"DNA Sequence","volume":"1 1","pages":"67 - 69"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74927462","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.3109/10425170109084464
A. S. Treglia, M. Gullì, C. Perrotta
A full-length cDNA encoding a novel small heat shock protein (HaHSP17.9) was isolated from a cDNA library of sunflower (Helianthus annuus cv. Gloriasol). The deduced amino acid sequence exhibited high degree homology to the class I cytosolic sHSPs from other plant species, and contained all the conserved regions characteristic of this class of proteins. Northern analyses showed that the transcript homologous to HaHSP17.9 accumulates during heat shock in suspension cultured cells and in the different parts of sunflower seedlings.
{"title":"Isolation and Characterisation of a cDNA for a Novel Small HSP from Sunflower Suspension Cell Cultures","authors":"A. S. Treglia, M. Gullì, C. Perrotta","doi":"10.3109/10425170109084464","DOIUrl":"https://doi.org/10.3109/10425170109084464","url":null,"abstract":"A full-length cDNA encoding a novel small heat shock protein (HaHSP17.9) was isolated from a cDNA library of sunflower (Helianthus annuus cv. Gloriasol). The deduced amino acid sequence exhibited high degree homology to the class I cytosolic sHSPs from other plant species, and contained all the conserved regions characteristic of this class of proteins. Northern analyses showed that the transcript homologous to HaHSP17.9 accumulates during heat shock in suspension cultured cells and in the different parts of sunflower seedlings.","PeriodicalId":11381,"journal":{"name":"DNA Sequence","volume":"121 1","pages":"397 - 400"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77015758","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.3109/10425170109042048
J. Morlon‐Guyot, F. Mucciolo-Roux, R. R. Sanoja, J. Guyot
Primers and probes were established from the sequences of the α-amylase genes (amyA) of L. amylovorus CIP 102989 and of L. plantarum A6 (Giraud and Cuny 1997). They were successfully used for the detection of the amy A gene in L. manihotivorans strain LMG 18010T and a 2842 bp region, containing the entire gene (2706 bp) with its putative promoter has been sequenced. More than 98% nucleotide sequence identities was found with L. amylovorus and L. plantarum amyA genes. The deduced amino acid sequence shares more than 96% amino acid sequence identities with L. amylovorus and L. plantarum ct-amy-lases, and also 65% and 59% identities with the cc-amy-lases of B. subtilis and S. bovis, respectively. The 3′ terminal part of L. manihotivorans LMG 18010TamyA gene contained four repeated sequences (SRU). The amyA genes of the three lactobacilli species differed mainly in the number of SRU and in the size of the flanking regions of the SRU.
{"title":"Characterization of the L. manihotivorans α-Amylase Gene","authors":"J. Morlon‐Guyot, F. Mucciolo-Roux, R. R. Sanoja, J. Guyot","doi":"10.3109/10425170109042048","DOIUrl":"https://doi.org/10.3109/10425170109042048","url":null,"abstract":"Primers and probes were established from the sequences of the α-amylase genes (amyA) of L. amylovorus CIP 102989 and of L. plantarum A6 (Giraud and Cuny 1997). They were successfully used for the detection of the amy A gene in L. manihotivorans strain LMG 18010T and a 2842 bp region, containing the entire gene (2706 bp) with its putative promoter has been sequenced. More than 98% nucleotide sequence identities was found with L. amylovorus and L. plantarum amyA genes. The deduced amino acid sequence shares more than 96% amino acid sequence identities with L. amylovorus and L. plantarum ct-amy-lases, and also 65% and 59% identities with the cc-amy-lases of B. subtilis and S. bovis, respectively. The 3′ terminal part of L. manihotivorans LMG 18010TamyA gene contained four repeated sequences (SRU). The amyA genes of the three lactobacilli species differed mainly in the number of SRU and in the size of the flanking regions of the SRU.","PeriodicalId":11381,"journal":{"name":"DNA Sequence","volume":"19 1","pages":"27 - 37"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81964313","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.3109/10425170109084454
Yoshinori Takahashi, K. Kako, K. Ohmura, K. Tsumori, Y. Ohmasa, Shin-ichi Kashiwabara, T. Baba, Eisuke Munekatat
Cox17p was first cloned as a cytoplasmic copper chaperone from yeast mutant and recent works suggested the existence of mammalian homologues. Previous report has shown that a gel filtration fraction of heart extract containing porcine Coxl7p peptide promoted the survival of NIH3T3 fibroblast cells. In the present study, we first cloned DNA fragments of the mouse COX17 gene. The mouse COX17 spans ∼6kb and consists of three exons. It was mapped to the center of chromosome 16, using a radiation hybrid-mapping panel. The major transcription start site is 80 bp upstream of the ATG initiation codon as determined by rapid amplification of cDNA ends (5′-RACE) analysis. Two potential polyadenylation sites are 3233 and 3293 bp downstream of the termination codon, respectively. Transient transfection of reporter plasmids containing portions of the mouse COX17 5′-flanking region into AtT-20 and NIH3T3 cells allowed the localization of the essential promoter to a 0.8 kb region upstream of the transcription starting site. Furthermore, the transfected luciferase activity was much higher in AtT-20 than NIH3T3. According to sequence analysis of the –0.8 kb 5′-flanking region, GC rich segments including consensus sequences for binding of the transcription factor Sp1, but no The TATAICAAT boxes, exist in the region of the transcription start site. Besides the GC box, binding sites for NRF-1 and 2 known as specific transcription factors for COX subunits are also localized around the transcription starting site.
{"title":"Genomic Structure of Mouse Copper Chaperone, 00×17","authors":"Yoshinori Takahashi, K. Kako, K. Ohmura, K. Tsumori, Y. Ohmasa, Shin-ichi Kashiwabara, T. Baba, Eisuke Munekatat","doi":"10.3109/10425170109084454","DOIUrl":"https://doi.org/10.3109/10425170109084454","url":null,"abstract":"Cox17p was first cloned as a cytoplasmic copper chaperone from yeast mutant and recent works suggested the existence of mammalian homologues. Previous report has shown that a gel filtration fraction of heart extract containing porcine Coxl7p peptide promoted the survival of NIH3T3 fibroblast cells. In the present study, we first cloned DNA fragments of the mouse COX17 gene. The mouse COX17 spans ∼6kb and consists of three exons. It was mapped to the center of chromosome 16, using a radiation hybrid-mapping panel. The major transcription start site is 80 bp upstream of the ATG initiation codon as determined by rapid amplification of cDNA ends (5′-RACE) analysis. Two potential polyadenylation sites are 3233 and 3293 bp downstream of the termination codon, respectively. Transient transfection of reporter plasmids containing portions of the mouse COX17 5′-flanking region into AtT-20 and NIH3T3 cells allowed the localization of the essential promoter to a 0.8 kb region upstream of the transcription starting site. Furthermore, the transfected luciferase activity was much higher in AtT-20 than NIH3T3. According to sequence analysis of the –0.8 kb 5′-flanking region, GC rich segments including consensus sequences for binding of the transcription factor Sp1, but no The TATAICAAT boxes, exist in the region of the transcription start site. Besides the GC box, binding sites for NRF-1 and 2 known as specific transcription factors for COX subunits are also localized around the transcription starting site.","PeriodicalId":11381,"journal":{"name":"DNA Sequence","volume":"19 1","pages":"305 - 318"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85097925","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2001-01-01DOI: 10.3109/10425170109084471
S. Hong, W. Yoo, R. Putnak, K. Eckels, H. Rho, Soo‐Ok Kim
Live attenuated Japanese encephalitis (JE) virus SA14-14-2 produced in primary dog kidney cells (PDK) was adapted to Vero cells. In an effort to gain insight into the molecular basis of the biological characteristics of the SA14-14-2(Vero) strain, the 1500 nucleotide sequence encoding the envelope (E) gene which possesses major neutralizing epitopes was determined and compared with the sequences of two other attenuated JE virus strains, SA14-14-2(PHK) and SA14-14-2(PDK). The amino acid sequence of the C-terminal region (a.a. 280-500) was found to be identical for all three strains, while the N-terminal region (a.a. 1-279) shows sequence variation. The distribution of mutations in the N-terminal region was nearly the same among the three attenuated strains, suggesting that the N-terminal sequences might be related with virus-host cell specificity. However, it was found that Lys and Val (a.a. 138 and 176, respectively), known to be responsible for attenuation, are still conserved in SA14-14-2(Vero). Animal testing showed that SA14-14-2(Vero) has an attenuation phenotype similar to that of the parent s SA14-14-2(PDK) strain in mice.
{"title":"Nucleotide Sequence of Envelope Protein of Japanese Encephalitis Virus SA14-14-2 Adapted to Vero Cells","authors":"S. Hong, W. Yoo, R. Putnak, K. Eckels, H. Rho, Soo‐Ok Kim","doi":"10.3109/10425170109084471","DOIUrl":"https://doi.org/10.3109/10425170109084471","url":null,"abstract":"Live attenuated Japanese encephalitis (JE) virus SA14-14-2 produced in primary dog kidney cells (PDK) was adapted to Vero cells. In an effort to gain insight into the molecular basis of the biological characteristics of the SA14-14-2(Vero) strain, the 1500 nucleotide sequence encoding the envelope (E) gene which possesses major neutralizing epitopes was determined and compared with the sequences of two other attenuated JE virus strains, SA14-14-2(PHK) and SA14-14-2(PDK). The amino acid sequence of the C-terminal region (a.a. 280-500) was found to be identical for all three strains, while the N-terminal region (a.a. 1-279) shows sequence variation. The distribution of mutations in the N-terminal region was nearly the same among the three attenuated strains, suggesting that the N-terminal sequences might be related with virus-host cell specificity. However, it was found that Lys and Val (a.a. 138 and 176, respectively), known to be responsible for attenuation, are still conserved in SA14-14-2(Vero). Animal testing showed that SA14-14-2(Vero) has an attenuation phenotype similar to that of the parent s SA14-14-2(PDK) strain in mice.","PeriodicalId":11381,"journal":{"name":"DNA Sequence","volume":"17 1","pages":"437 - 442"},"PeriodicalIF":0.0,"publicationDate":"2001-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82640256","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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