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Molecular cloning of the Candida maltosa ADE1 gene. 总被引:1,自引:0,他引:1
The structural gene (ADE1) encoding phosphoribosyl-aminoimidazole-succinocarboxamide synthetase (SAICAR synthetase; EC 6.3.2.6) in Candida maltosa has been isolated by functional complementation of an ade1 strain of Saccharomyces cerevisiae. The gene was localized on a 2.5-kb BamHI DNA fragment. Nucleotide sequence analysis of the cloned gene has revealed an open reading frame encoding a protein (SAICAR synthetase) with an Mr of 32,751. The codon bias index, 0.68, indicates that the ADE1 gene is a moderately highly expressed gene. The cloned gene shows 63.5% nt identity and 65.2% deduced amino acid identity with the S. cerevisiae ADE1 gene which encodes the same enzymatic activity. The gene may be used as a convenient genetic marker for construction of a new host-vector system for C. maltosa. 相似文献
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Oocytes are recognized as a source of regulatory molecules that influence follicular development through an array of actions on granulosa cells. Recently, more and more hormones and signaling molecules were identified during follicular developmental processes; however, the details about their functions are still unclear. During efforts to clone follicular development-related genes, we isolated a cDNA fragment by DDRT-PCR. To obtain cDNA 5'- and 3'-end sequences, we screened a mouse ovarian cDNA library. After screening the library, an open reading frame of 2,994 bp for the new gene (Oocyte-G1), which encodes a 997-residue protein, was cloned. Northern blot analysis revealed the presence of approximately 3.6 kb Oocyte-G1 mRNA in ovary, lung, kidney, testis and brain. Northern analysis of RNA from ovaries in vivo showed that Oocyte-G1 was weakly expressed on day 5 and at a moderate level on day 10. Thereafter, on day 15 or in adults (day 40), there was an increase in expression, followed by a decline in ovaries on day 20 or older (day 120). Furthermore, we studied the Ooctye-G1 protein by using the antiserum against a peptide sequence unique to this gene in Western blotting and immunolocalization. The antiserum recognized a prominent band of approximately 110 kDa in immunoblots and signals were dispersed in oocytes and some cumulus granulosa cells. Our results suggest the potential role of Oocyte-G1 in ovarian follicular development. 相似文献
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DNA fragments complementing theenvC mutation could be isolated by cloning chromosomal DNA in the vector pUH84. When the frequencies of transformation and the frequencies of restoring theenvC
+ phenotype were compared, the high copy number hybrid plasmids complemented with a frequency of 10–5. After subcloning theenvC-complementing DNA fragment into the low copy number plasmid pLG339, efficient complementation was achieved by spontaneous integration of the IS2 element ofEscherichia coli. By nucleotide sequence analysis, a potential promoter, a ribosome-binding site, and an unidentified reading frame were detected in the respective DNA fragment. 相似文献
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N S Neznanov B M Troianovski? A L Gartel' I V Makarova K A Bendukidze 《Molekuliarnaia genetika, mikrobiologiia i virusologiia》1987,(7):14-15
Isolation of genomic clones containing the human insulin gene by screening the human genomic library for this gene using the cDNA rat insulin probe is reported. The analysis of promoter-enhancer region and exons sequences has revealed their identity to analogous sequences determined earlier. 相似文献
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Molecular cloning of the mouse angiotensinogen gene 总被引:7,自引:0,他引:7
We describe here the cloning, restriction mapping, and sequencing of the mouse angiotensinogen gene. The 5' flanking region contains consensus sequences for several hormone-responsive elements and viral-like enhancers within 750 bp of the cap site. The deduced amino acid sequence shows 87% identity with rat angiotensinogen, but there is a discrepancy in the number of cysteine residues in the mature protein among rat (n = 3), human (n = 4), and mouse (n = 4). Because angiotensinogen is homologous to other members of the serine protease inhibitor family, we aligned the putative reactive center of angiotensinogens from various species. This alignment shows that the inhibitor site in human angiotensinogen is different from its rodent counterpart, but the role of this sequence divergence in the pathogenesis of human disease remains to be established. 相似文献
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K Hori Y Yamamoto T Minetoki T Kurotsu M Kanda S Miura K Okamura J Furuyama Y Saito 《Journal of biochemistry》1989,106(4):639-645
The entire gene for gramicidin S synthetase 1 (GS 1) was cloned into the plasmid vector pUC18, and the nucleotide sequences of the GS 1 gene and its flanking region were determined. The full-length clone was 4,539 base pairs long and had an open reading frame of 3,294 nucleotides coding for 1,098 amino acids. The calculated molecular weight of 123,474 agreed with the apparent molecular weight of 120,000 found in SDS-PAGE of GS 1 from B. brevis. The nucleotide sequence of GS 1 gene was highly homologous to that of tyrocidine synthetase 1. The overall similarity between the deduced amino acid sequences of the two genes was 57.5%. The gene product of clone GS309 was easily purified to an essentially homogeneous state by ammonium sulfate fractionation followed by DEAE-Sepharose CL-6B, Ultrogel AcA-34, and second DEAE-Sepharose CL-6B column chromatography. The purified protein catalyzed the D-phenylalanine-dependent ATP-32PPi exchange reaction which is specific for GS 1 activity, and the specific activity of the purified product was nearly the same as the purified GS 1 from B. brevis. The product also showed a weak phenylalanine racemase activity. 相似文献
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Tsutsumi S Inoue H Sakamoto Y Mizuta K Kamata N Itakura M 《Biochemical and biophysical research communications》2005,331(4):1099-1106
Mutations in the GDD1 gene cause gnathodiaphyseal dysplasia, a rare human skeletal syndrome with autosomal dominant inheritance. The biochemical function(s) of GDD1 protein and the molecular pathophysiology of GDD1 mutations leading to GDD have not yet been elucidated. In this study, we characterized the complete cDNA sequence and genomic organization of the mouse GDD1 gene. Analysis of GDD1 mRNA revealed a complex alternative splicing pattern, involving five exons of the GDD1 gene. GDD1 isoforms lacking conserved amino acids at the N-terminus cytoplasmic tails, and with changes in transmembrane topology, are presumably associated with changes in protein functions and subcellular localizations of GDD1. We found GDD1 expression to be up-regulated during the course of myogenic differentiation in the murine pluripotent mesenchymal precursor cell line C2C12, whereas its expression was diminished during osteoblastic differentiation. These observations suggest diverse cellular roles of GDD1 protein. 相似文献
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Molecular cloning and characterization of an activated human c-raf-1 gene. 总被引:6,自引:2,他引:4 下载免费PDF全文
M Fukui T Yamamoto S Kawai F Mitsunobu K Toyoshima 《Molecular and cellular biology》1987,7(5):1776-1781
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细胞周期蛋白cyclin G1(CCNG1)是一个重要的细胞周期调控因子,参与哺乳动物颗粒细胞增殖、卵母细胞成熟等繁殖生物学过程,但其在绵羊中鲜有报道。为了研究CCNG1基因对绵羊发情调控以及季节性繁殖的影响,文章对绵羊CCNG1基因进行了克隆和组织表达谱分析,利用Real-time PCR对该基因在多浪羊(常年发情)与中国美利奴羊(季节性繁殖)发情周期不同阶段性腺轴组织的表达变化进行了实时检测。获得了绵羊CCNG1基因部分cDNA序列,其中编码区全长885 bp,编码294个氨基酸。CCNG1蛋白结构经预测存在多个磷酸化位点和蛋白激酶C磷酸化位点。CCNG1基因在所检测绵羊各组织中均有表达,但在卵巢与肾脏中为高丰度表达;CCNG1在不同绵羊品种发情周期不同阶段性腺轴组织的表达变化规律基本一致,卵巢、子宫、松果体、垂体均是在发情期达到峰值。但是在发情期和发情后期卵巢CCNG1的表达量存在显著的品种间差异(P<0.01)。研究结果表明,CCNG1可能通过参与卵泡的生长发育继而达到对绵羊发情和季节性繁殖的调控。 相似文献
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Yasunobu Kano Setsuo Yoshino Morimasa Wada Kazushige Yokoyama Masahiro Nobuhara Fumio Imamoto 《Molecular & general genetics : MGG》1985,201(2):360-362
Summary The Escherichia coli HU-1 was cloned by use of mixed synthetic oligonucleotides (17-mer) predicted from a portion of its amino acid sequence. The amino acid sequence of the HU-1 protein deduced from the nucleotide sequence is in good agreement with the published sequence. The nucleotide sequence has a possible promoter and a typical ribosomal binding site upstream from the translational initiation codon (GUG) of the HU-1 gene. 相似文献
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The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have
been assigned the accession number L50534 相似文献
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Norio Kurosawa Kazunori Kondo Naohiro Kimura Tomio Ikeda Yasuzo Tsukada 《Neurochemical research》1994,19(5):575-580
Birds have several advantages in the study of memory formation, as imprinting and passive avoidance behaviors in chick are often used as model systems. However, the primary structure of the bird N-methyl-d-aspartate (NMDA) responsive glutamate receptor, which is assumed to play a critical role in memory formation, has not been determined. In this report we describe the cDNA cloning of a subunit of NMDA receptors (NMDA-R1) from duck and analysis of its structure and distribution in the brain. The N-terminal 898 amino acids of the NMDA-R1 were well conserved between duck and mammals, but the homology was completely lost in the C-terminus. In situ hybridization showed that the duck NMDA-R1 gene was expressed throughout the brain as it is in mammals.Special issue dedicated to Dr. Bernard W. Agranoff. 相似文献