المؤلفون: Sotiria Boukouvala, Edith Sim

المصدر: Basic Toxicology. 96:343-351

مصطلحات موضوعية: Gene isoform, Transcription, Genetic, Arylamine N-Acetyltransferase, Molecular Sequence Data, Biology, Polyadenylation, Toxicology, Exon, Complementary DNA, Humans, Coding region, Gene, Expressed Sequence Tags, Pharmacology, Genetics, Genomic Library, Expressed sequence tag, Reverse Transcriptase Polymerase Chain Reaction, Alternative splicing, Intron, Chromosome Mapping, Exons, General Medicine, Molecular biology, Introns, Isoenzymes, Alternative Splicing, Genes, Sequence Alignment, Chromosomes, Human, Pair 8

الوصف: Arylamine N-acetyltransferases are polymorphic drug-metabolising enzymes. The human isoforms, NAT1 and NAT2, are encoded by two genes with intronless coding regions. Human NAT1 protein is found in many tissues, unlike NAT2 which is present predominantly in the intestine and liver. We describe the exon-intron structure of the human NAT genes by analysing data from genomic databases. Comparison of expressed sequence tags, matching NAT gene sequences, with the sequence of human chromosome 8 implied the presence of 8 non-coding exons located 51.5, 51.4, 12.3, 11.9, 10.8, 9.6, 5.2 and 2.6 kb upstream of the single coding exon of the NAT1 gene. A number of expressed sequence tags also indicated transcription initiation from the upstream region adjacent to the NAT1 coding exon, consistent with earlier studies. The NAT2 gene consists of one previously described non-coding and one coding exon, located 8.6 kb apart. These findings were also confirmed by RT-PCR, using cDNA from heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. Alternatively spliced NAT1 transcripts were found in all tissues. Transcription of the NAT2 gene was also detected in these tissues and was demonstrated to start either from the non-coding exon or from immediately upstream of the coding exon. Comparison of the RT-PCR products provided an initial estimate of the relative amounts of the different NAT transcripts expressed in each tissue. Finally, both expressed sequence tag analysis and RT-PCR demonstrated the presence of two differentially utilised polyadenylation signals for NAT1 and NAT2, located about 0.2 and 0.3 kb downstream of the coding region of each gene.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2556fe0284a9ec17e11f5ad2f56301d8Test
https://doi.org/10.1111/j.1742-7843.2005.pto_02.xTest

المؤلفون: Sayaka Itoh, Hideomi Amano, Yasutoshi Yoshiura, Eri Inagaki, Makoto Kakinuma, Daniel A. Coury

المصدر: Gene. 334:145-155

مصطلحات موضوعية: Photoperiod, Molecular Sequence Data, Gene Dosage, Gene Expression, Chlorophyta, Ulva, Complementary DNA, Gene expression, Genetics, Coding region, Amino Acid Sequence, RNA, Messenger, Gene, Base Sequence, Sequence Homology, Amino Acid, biology, cDNA library, DNA, Sequence Analysis, DNA, General Medicine, Blotting, Northern, biology.organism_classification, Molecular biology, Actins, Culture Media, Open reading frame, Ulvales, Genes, Mutation, Sequence Alignment

الوصف: We constructed a cDNA library from sterile Ulva pertusa (Ulvales, Chlorophyta), and isolated and characterized a full-length cDNA clone encoding actin. The actin (ACT) cDNA consisted of 1487 nucleotides (nt) and had an open reading frame (ORF) encoding a polypeptide of 377 amino acid (AA) residues. The ACT gene had one intron in the 5'-untranslated region and three introns in the coding region. Transcription started 26 nt downstream of the putative TATA box. A potential polyadenylation signal, TGTAG, was located 100 nt downstream of the terminator codon, TAG. Amino acid alignment with actins from various algae and land plants showed that sterile U. pertusa actin was more similar to actins from Chlorophyta, Phaeophyta, Euglenophyta, and higher plants (over 76.9%) than to actins from Rhodophyta. Southern blot analysis indicated that the sterile U. pertusa genome has only a single actin-encoding gene. Thalli grown on a 12D/12L photoperiod increased in surface area some two-fold over 24 h regardless of the nutritional conditions. The growth rate of thalli during the light period was significantly higher than that during the dark period. Northern hybridization indicated that the expression of actin mRNA was induced and repressed by the light and dark treatments, respectively. These results suggest that the U. pertusa cell division cycle has a periodicity of 24 h and that the ACT gene is highly transcribed during cell growth and development in the light period.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::6af2935abbfdd1a498ac5f8c3a968519Test
https://doi.org/10.1016/j.gene.2004.03.029Test

المؤلفون: Lei Xu, Yi Yu, Bingwei Wang, Zhihua Li, Jiawei Zhou, Julian Goggi

المصدر: Gene. 331:149-157

مصطلحات موضوعية: alpha Karyopherins, Untranslated region, DNA, Complementary, Sequence analysis, Molecular Sequence Data, Gene Expression, Biology, Rats, Sprague-Dawley, Hydroxydopamines, Exon, Complementary DNA, Genetics, Importin-alpha, Animals, Coding region, 3' Flanking Region, Amino Acid Sequence, RNA, Messenger, Northern blot, Cloning, Molecular, Karyopherin, chemistry.chemical_classification, Base Sequence, Sequence Homology, Amino Acid, Gene Expression Profiling, Exons, Sequence Analysis, DNA, General Medicine, Molecular biology, Corpus Striatum, Introns, Rats, Genes, chemistry, Female, Databases, Nucleic Acid, Sequence Alignment

الوصف: Dopamine denervation in the striata of patients with Parkinson's disease (PD) leads to changes in neural plasticity. However, the mechanisms leading to the changes are still poorly understood. In an effort to study the molecular events in the denervated striatum, we identified and cloned rat karyopherin α1 (KPNA1), a member of the importin/karyopherin α (KPNA) family. DNA sequence analysis revealed that the full-length cDNA, encoding rat KPNA1, was 4975 bp with a short 5′-untranslated region (UTR) of 70 bp, a putative coding sequence of 1617 bp, and an unusually long 3′-UTR of 3266 bp. The gene shared a high degree of similarity with its mouse and human homologs at both cDNA and protein levels. By computational analysis of its genomic sequence, the transcription unit was shown to span a 44-kb region and consist of 13 exons varying in size from 89 (6th exon) to 3454 bp (13th exon), and 12 introns varying in size from 0.3 to 8.9 kb. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis demonstrated that KPNA1 transcript existed in various adult tissues. Both Northern blot and semi-quantitative RT-PCR analysis showed that the expression level of KPNA1 mRNA was altered in the denervated striatum post-lesion in a time-dependent manner, reaching the maximum at 2 weeks post-lesion. Our results suggest involvement of KPNA1 in the striatal responses to denervation following 6-hydroxydopamine (6-OHDA)-induced lesion.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::9f809e6e47e585f53a217d329b48bf54Test
https://doi.org/10.1016/j.gene.2004.02.009Test

المؤلفون: Siu-Ming Chan, Jian-Guo He, Shirley H.K. Tiu, Billy K. C. Chow, L. Scott Quackenbush, Wing-Sze Tsang

المصدر: Gene. 303:99-109

مصطلحات موضوعية: DNA, Complementary, animal structures, Sequence analysis, Molecular Sequence Data, Gene Expression, Vitellogenins, Vitellogenin, Penaeidae, Rapid amplification of cDNA ends, Sequence Homology, Nucleic Acid, Complementary DNA, Genetics, Animals, Coding region, Amino Acid Sequence, Cloning, Molecular, Gene, Phylogeny, Base Sequence, biology, Ovary, Exons, Sequence Analysis, DNA, General Medicine, Molecular biology, Introns, Shrimp, Genes, Multigene Family, biology.protein, Female, Digestive System

الوصف: Vitellogenin is the major egg yolk protein synthesized in female shrimp during gonad maturation. Although there are several reports for the cloning of vitellogenin complementary DNA (cDNA) in different crustaceans, little is known of the gene organization of this protein. This study reports the first cloning and characterization of a full-length gene encoding the vitellogenin precursor from the shrimp Metapenaeus ensis. By genomic DNA library screening, six different lambda clones were isolated using shrimp partial gene sequence as probe. Initial DNA sequence determination revealed that these clones are derived from different genes with coding sequence similar to other crustacean vitellogenins. Two of these clones were used for further analysis. One of the lambda clones (lambda 3.3) carries most of the coding sequence that correspond to the M. ensis vitellogenin gene (MeVg1) and the other clone (lambda 8.3) carries a smaller portion of the coding sequence of a different vitellogenin gene (MeVg2). The lambda 3.3 clone was chosen for further characterization. To clone the remaining 5' end upstream promoter region, 5' untranslated region and the remaining coding sequence of MeVg1, a polymerase chain reaction (PCR)-based gene walking approach was used. Subsequently, a PCR clone with overlapping sequence identical to the genomic clone was obtained and the organization of MeVg1 gene was constructed. The MeVg1 gene consists of 15 exons and 14 introns spanning approximately 10 kb. Several potential cleavage sites were identified from the deduced vitellogenin precursor. Cleaving of the precursor in these sites would result in the production of several vitellogenin subunits. To clone the cDNA for the vitellogenin, 5' and 3' rapid amplification of cDNA ends was performed using ovary cDNA of the shrimp. A 4.4 kb 5' cDNA clone and a 4 kb 3' end cDNA clone were isolated. The size of the reconstructed cDNA for M. ensis Vg is 7.97 kb and consists of the longest open reading frame of 7776 bp. Unlike the vitellogenin precursor of most insects and vertebrates, the deduced vitellogenin precursor lacks the polyserine domain important for receptor-mediated endocytosis. Phylogenetic analysis revealed a closer relationship of the MeVg1 with other crustacean vitellogenins but distantly related to other invertebrate and vertebrate vitellogenins. By reverse transcription-PCR, we have demonstrated that the shrimp MeVg1 gene is expressed only in the ovary and hepatopancreas while the MeVg2 gene is expressed exclusively in the hepatopancreas. In conclusion, the shrimp ovary also contribute significantly in the production of vitellogenin at transcription level and the gene organization of the shrimp protein may provide an insight in the evolution of this group of important proteins.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c448ce2c6fc762bbd52afc843f5cf431Test
https://doi.org/10.1016/s0378-1119Test(02)01139-3

المؤلفون: Robert A. Peterfreund, James F. Gusella, Mia MacCollin, J S Fink

المصدر: Journal of Neurochemistry. 66:362-368

مصطلحات موضوعية: DNA, Complementary, Chromosomes, Human, Pair 22, Molecular Sequence Data, Gene Expression, Nerve Tissue Proteins, Biology, Biochemistry, Cellular and Molecular Neuroscience, Sequence Homology, Nucleic Acid, Complementary DNA, Gene expression, Humans, Coding region, Amino Acid Sequence, RNA, Messenger, Northern blot, Lung, Gene, Southern blot, Brain Chemistry, Base Sequence, cDNA library, Receptors, Purinergic P1, Molecular biology, Adenosine receptor, Introns, Genes, Organ Specificity

الوصف: The actions of the neurotransmitter adenosine are mediated by a family of high-affinity, G protein-coupled receptors. We have characterized the gene for the human A2a subtype of adenosine receptor (hA2aR) and determined levels of A2aR mRNA in human brain regions and nonneural tissues. Human genomic Southern blot analysis demonstrates the presence of a single gene encoding the hA2aR located on chromosome 22. Two overlapping cosmids containing the hA2aR gene were isolated from a chromosome 22 library and characterized. Southern blot and sequence analyses demonstrate that the hA2aR gene spans approximately 9-10 kb with a single intron interrupting the coding sequence between the regions encoding transmembrane domains III and IV. The sequence of the hA2aR gene diverged from the reported cDNA structure in the 5' untranslated region. This divergence appears to result from an artifact in the construction of the original cDNA library. By northern blot analysis, high expression of the hA2aR gene was identified in the caudate nucleus with low levels of expression in other brain regions. High expression was also seen in immune tissues; lesser A2aR expression was detected in heart and lung. The gene for the A2a subtype of receptor for the neurotransmitter adenosine falls in the class of intron containing G protein-coupled receptor genes. Expression in the basal ganglia is consistent with a role for the hA2aR in motor control. Activation of the A2aR may also regulate immune responses and cardiopulmonary function.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1af4c5f6a353f7c63ceb39d0d646c337Test
https://doi.org/10.1046/j.1471-4159.1996.66010362.xTest

المؤلفون: Paula D. Ladd, Diana L. Carlone, David G. Skalnik, Suzanne R.L. Hart

المصدر: Gene. 295:71-77

مصطلحات موضوعية: DNA, Complementary, Molecular Sequence Data, Protein domain, Gene Expression, Mice, Inbred Strains, Biology, Cell Line, Mice, Exon, Complementary DNA, Genetics, Animals, Coding region, Amino Acid Sequence, Cloning, Molecular, Gene, Cloning, Mice, Inbred C3H, Binding Sites, Base Sequence, Sequence Homology, Amino Acid, Nucleic acid sequence, DNA, Exons, Sequence Analysis, DNA, General Medicine, Molecular biology, Introns, DNA-Binding Proteins, Genes, CpG site, CpG Islands

الوصف: Human CpG binding protein (CGBP) is a ubiquitously-expressed transcriptional activator that binds specifically to unmethylated CpG motifs. Several protein domains have been identified within CGBP including two plant homeodomains (PHD), acidic and basic regions, a coiled-coil domain, as well as a CXXC DNA-binding domain. The global function of CGBP remains unclear, although failure to express CGBP results in embryonic lethality in mice. This study reports the identification and characterization of the murine CGBP gene locus. A 2509 bp murine CGBP cDNA was cloned and nucleotide sequence determined. Comparison of the mouse and human CGBP sequences revealed 86% identity at the nucleotide level and 96% identity at the amino acid level. Examination of the deduced translation product revealed that the PHD, CXXC, coiled-coil, and basic domains are identical between mouse and human, while the acidic region exhibits approximately 90% identity with its human counterpart. A single murine CGBP transcript of approximately 2.6 kb was detected in a wide variety of adult tissues as well as embryonic stem cells. Analysis of the mouse gene locus revealed a relatively small gene spanning approximately 5 kb and comprised of 15 exons. Examination of the human CGBP gene showed a similar size and structure with identical intronic splice sites. In contrast to the human CGBP gene, which is located 800 bp upstream of the MBD1 gene, analysis of the murine CGBP gene locus failed to detect the murine MBD1 gene within several kilobases of the CGBP coding region.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8fe9b56187e7b90e5eb8e2092eda0658Test
https://doi.org/10.1016/s0378-1119Test(02)00820-x

المؤلفون: Quan Zhen Li, Cong-Yi Wang, Wei Lian, Donghai Wu, Qing Guo Ruan, Sarah Eckenrode, Jin Da Shi, Thomas Kukar, Yunrong Gu, Jin-Xiong She, Abdoreza Davoodi-Semiromi

المصدر: Gene. 273:275-284

مصطلحات موضوعية: Male, Cytoplasm, DNA, Complementary, Recombinant Fusion Proteins, Green Fluorescent Proteins, Molecular Sequence Data, Gene Expression, Tubby protein, Molecular cloning, Biology, Cell Line, Mice, WD40 repeat, Complementary DNA, Diabetes Mellitus, Genetics, Animals, Humans, Coding region, Gene family, Tissue Distribution, Amino Acid Sequence, RNA, Messenger, Cloning, Molecular, Dinucleotide Repeats, Gene, Phylogeny, Adaptor Proteins, Signal Transducing, Polymorphism, Genetic, Sequence Homology, Amino Acid, Intracellular Signaling Peptides and Proteins, Chromosome Mapping, Proteins, Sequence Analysis, DNA, General Medicine, Blotting, Northern, Alternative Splicing, Luminescent Proteins, Genes, Chromosomes, Human, Pair 6, Female, Sequence Alignment, Nuclear localization sequence

الوصف: We report here the cloning and characterization of a novel gene belonging to the tubby superfamily proteins (TUSP) in mouse and human. The mouse Tusp cDNA is 9120 bp in length and encodes a deduced protein of 1547 amino acids, while the human TUSP gene is 11,127 bp and encodes a deduced protein of 1544 amino acids. The human and mouse genes are 87% identical for their nucleotide sequences and 85% identical for their amino acid sequences. The protein sequences of these genes are 40–48% identical to other tubby family proteins at the C-terminal conserved ‘tubby domain’. In addition, the TUSP proteins contain a tubby signature motif (FXGRVTQ), two bipartite nuclear localization signals (NLSs) at the C-terminal, two proline-rich regions, one WD40 repeat region and one suppressor of cytokines signaling domain. Transfection assay with green fluorescent protein-tagged TUSP expression constructs showed that the complete TUSP protein and the N-terminal portion of TUSP are localized in the cytoplasm but the C-terminal portion with the two NLSs produced distinct dots or spots localized in the cytoplasm. Northern blotting analysis showed that the major transcript with the complete coding sequence is expressed mainly in the brain, skeletal muscle, testis and kidney. Radiation hybrid mapping localized the mouse gene to chromosome 17q13 and the human TUSP gene to chromosome 6q25-q26 near the type 1 diabetes gene IDDM5 . However, association analysis in diabetic families with a polymorphic microsatellite marker did not show any evidence for association between TUSP and type 1 diabetes. The precise biological function of the tubby superfamily genes is still unknown; the highly conserved tubby domain in different species, however, suggests that these proteins must have fundamental biological functions in a wide range of multi-cellular organisms.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::28f4b225e9b7d5b29d8807295845bdaeTest
https://doi.org/10.1016/s0378-1119Test(01)00582-0

المؤلفون: Amy L. Tucker, Roberta C. Bogaev, Li Guo Jia, Cathy J. Palmer, Larry R. Jones, Yvonne M. Kobayashi, J. Randall Moorman, J. Paul Mounsey

المصدر: Gene. 271:69-79

مصطلحات موضوعية: Male, Untranslated region, DNA, Complementary, Molecular Sequence Data, Phospholemman, CAAT box, Gene Expression, Mice, Inbred Strains, Biology, Mice, Exon, Dogs, Cricetinae, Microsomes, Sequence Homology, Nucleic Acid, Complementary DNA, Genetics, Animals, Coding region, Tissue Distribution, Amino Acid Sequence, RNA, Messenger, Cloning, Molecular, Gene, Mice, Inbred BALB C, Base Sequence, Sequence Homology, Amino Acid, Myocardium, fungi, Alternative splicing, Gene Expression Regulation, Developmental, Membrane Proteins, DNA, Exons, Sequence Analysis, DNA, General Medicine, Blotting, Northern, Embryo, Mammalian, Phosphoproteins, Molecular biology, Introns, Rats, Alternative Splicing, Genes, Rabbits, Sequence Alignment

الوصف: Phospholemman (PLM) is a small transmembrane cardiac protein that is the major sarcolemmal substrate for phosphorylation in response to adrenergic stimulation. PLM likely plays a role in muscle contractility and cell volume regulation through its function as a channel or a channel regulator. We are the first to describe the structure of the PLM gene and to demonstrate PLM cDNA splice variants. We cloned the murine PLM cDNA and used it as a probe to isolate the gene from a 129/SvJ genomic library. The gene contains seven introns and eight exons. The coding sequence is interrupted by five introns; the 5' untranslated region by two. Using rapid amplification of 5' cDNA ends we identified transcription start sites and four splice variants of the 5' untranslated domain. There was no TATA box or CAAT box in the putative promoter regions. The gene has several stretches of dinucleotide repeats. The 3' untranslated domains of mouse PLM cDNA clones show sequence differences not accounted for by alternative splicing. Mouse PLM shares 93, 83 and 80% amino acid identity with rat, dog, and human PLMs, respectively. Tissue expression of murine PLM parallels that in other species, being highest in heart, skeletal muscle, and liver.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::05f3dc4607c8ae4181d1bfda817839b8Test
https://doi.org/10.1016/s0378-1119Test(01)00497-8

المؤلفون: Kirstin R.W. Matthews, Rick A. Wetsel

المصدر: The Journal of Immunology. 166:6196-6202

مصطلحات موضوعية: DNA, Complementary, Transcription, Genetic, Sequence analysis, Molecular Sequence Data, Immunology, Gene Dosage, Biology, Primer extension, Mice, Exon, Complementary DNA, Animals, Humans, Lysine Carboxypeptidase, Immunology and Allergy, Coding region, Amino Acid Sequence, Cloning, Molecular, Gene, Base Sequence, Intron, Chromosome Mapping, Blotting, Northern, Molecular biology, Mice, Inbred C57BL, Open reading frame, Genes, Liver, Cattle, 5' Untranslated Regions

الوصف: Carboxypeptidase N (CPN) is a plasma zinc metalloprotease comprised of two small subunits that have enzymatic activity, and two large subunits, which protect the enzyme from degradation. CPN cleaves the carboxyl-terminal amino acids arginine and lysine from biologically active peptides such as complement anaphylatoxins, kinins, and fibrinopeptides. To delineate the murine CPN small subunit coding region, gene structure, and chromosome location, cDNA and genomic clones were isolated, characterized, and used in Northern and fluorescence in situ hybridization analyses. The results from this study demonstrate that the murine CPN small subunit gene is a single copy gene of ∼29 kb that is transcribed in the liver into a 1793-bp mRNA with an open reading frame of 1371 nucleotides encoding 457 aa. The gene contains nine exons ranging in size from 455 bp (exon 1) to 100 bp (exon 7), and eight introns ranging in size from 6.2 kb (intron 2) to 1.4 kb (intron 4). All intron/exon junctions follow the normal consensus rule. The mouse CPN small subunit gene localized to chromosomal band 19D2, which is syntenic to human chromosome 10q23–25. Primer extension experiments using mouse liver mRNA indicate one major transcriptional initiation site and three minor sites. Sequence analysis of the 5′-flanking region indicated a TATA-less promoter and numerous transcription factor binding sites, which may confer liver-specific expression of the CPN small subunit gene.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::f58e21ed743c1d0ce6b1e2467c8317bdTest
https://doi.org/10.4049/jimmunol.166.10.6196Test

المؤلفون: Martine Yerle, Corinne Delville-Giraud, Sylvie Rival, Claire Rogel-Gaillard, Joe Attal, Louis-Marie Houdebine, Pascale Laffont

المساهمون: Unité biologie du développement et biotechnologie, École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Unité de recherche Génétique Biochimique et Cytogénétique (LGBC), Institut National de la Recherche Agronomique (INRA), Laboratoire de Génétique Cellulaire (LGC), Institut National de la Recherche Agronomique (INRA)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Laboratoire de radiobiologie et d'étude du génome (LREG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de la Recherche Agronomique (INRA), École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT), Institut National de la Recherche Agronomique (INRA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

المصدر: HAL
ResearcherID
Gene
Gene, Elsevier, 2001, 267, pp.37-47
Gene, 2001, 267, pp.37-47

مصطلحات موضوعية: Transcription, Genetic, Swine, [SDV]Life Sciences [q-bio], Molecular Sequence Data, Gene Expression, Biology, Cell Line, Mice, 03 medical and health sciences, Exon, Mammary Glands, Animal, Plasmid, Sequence Homology, Nucleic Acid, Complementary DNA, Genetics, Animals, Lactation, Coding region, Amino Acid Sequence, Cloning, Molecular, Gene, In Situ Hybridization, Fluorescence, 030304 developmental biology, 0303 health sciences, Bacterial artificial chromosome, Base Sequence, 030302 biochemistry & molecular biology, Chromosome Mapping, DNA, Sequence Analysis, DNA, General Medicine, BIOLOGIE MOLECULAIRE, Milk Proteins, Molecular biology, Chromosome Banding, LIGNE CELLULAIRE, Genes, Regulatory sequence, biology.protein, RNA, Rabbits, Whey Acidic Protein

الوصف: The whey acidic protein (WAP) is the major whey protein of rodent, rabbit and camel. Recently, it was identified in the milk of swine ( Simpson et al., 1998 . J. Mol. Endocrinol. 20, 27–35). In this paper, the cloning of the pig WAP cDNA and of bacterial artificial chromosome (BAC) construct containing the entire porcine WAP gene is reported. The comparison of the coding sequence of the pig WAP gene to rodent or lagomorph WAP sequence already published demonstrated that only exon sequences are partially conserved. The porcine WAP gene was localized on the subtelomeric region of the chromosome 18. The estimation of the expression of the swine WAP gene in the mammary gland from lactating animals revealed a high level of expression. In order to compare the expression level of the porcine WAP gene from the large genomic fragment which contained 70 kb downstream and 50 kb upstream the pig WAP gene or the smaller one (1 kb downstream and 2.4 kb upstream), these two genomic fragments were transfected in HC11 cell line. The BAC construct was expressed 15 times higher than the plasmid when reported to the integrated copy number. This report suggests that the HC11 cell line is a useful tool to identify the regulatory sequences of milk protein genes.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::22231779789d6c04495d367fff3c329aTest
https://doi.org/10.1016/s0378-1119Test(01)00388-2