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  • 1
    ISSN: 0378-1119
    Keywords: Gene isolation ; homologies ; nucleotide and amino acid sequences
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1438-2199
    Keywords: Amino acids ; Potassium channel ; Yeast ; Transcription regulation ; Spasmolytic and hypotensive drugs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Living cells control their electrical responsiveness by regulating the quality and quantity of channels expressed in the plasma membrane. Regulation of transcription of the voltage-gated ion channels is an important part of the molecular basis of cell energization. However, the factors which control the expression of channels are not well understood. We studied the effect on the transcription of the voltage-gated K+ channel in the yeastSchizosaccharomyces pombe of cations, pH, and therapeutic spasmolytic and hypotensive agents with different mechanisms of action, including accumulation of intracellular cAMP. A highly specific 122 bp domain of the K+ channel between S5 and H5 with a 55% homology with Dros shab and mbk3 was amplified by nested PCR from chromosomal DNAS. pombe. Northern blot revealed a 1.8kb transcript. mRNA dot-blot and RNase-protected analysis revealed factors altering the K+ channel transcription.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Amino acids 17 (1999), S. 301-313 
    ISSN: 1438-2199
    Keywords: Amino acids ; Taurine ; Transporter ; Rat ; Brain ; Heart
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary In pro- and eucaryotic life, cellular and subcellular compartments are separated by membranes and the regulated and selective passage of specific molecules across these membranes is a basic and highly conserved principle. We were interested whether taurine, a naturally occuring amino acid, would be able to induce or suppress expression of transporters with the Rationale that taurine was shown to detoxify a series of endogenous toxins and xenobiotics of various chemically non-related structures. For this purpose we used a gene hunting technique, subtractive hybridization, subtracting mRNAs of taurine-treated rat brain and heart from untreated controls. Subtracted mRNAs were then converted to cDNAs, amplified, sequenced and identified by gene bank data. We found five transporter transcripts, the phosphonate transport ATPase PHNC, multidrug transporter homolog MTH104, protein-exportmembrane protein SECD, oligopeptide transporters oppA and oppD, in the brain and two: ABC-transporter BRAF-2 and cation-transport ATPase PACS, in the heart. Homologies of the sequences found were in any case 〉50% thus permitting the identification of transporters with high probability. The biological meaning could be that a naturally occuring amino acid, taurine, modulates complex transport systems. The most prominent finding is the upregulation of a multidrug transporter transcript, explaining a mechanism for the nonselective detoxifying action of taurine.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1438-2199
    Keywords: Amino acids ; Taurine ; Osmoregulation ; Rat ; Osmolarity sensor protein ENVZ
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Although the involvement of taurine in osmoregulation is well-documented and widely accepted, no detailed mechanism for this function has been reported so far. We used subtractive hybridization to study mRNA steady state levels of genes up- or downregulated by taurine. Rats were fed taurine 100mg/kg body weight per day for a period of three days and hearts (total ventricular tissue) of experimental animals and controls were pooled and used for mRNA extraction. mRNAs from two groups were used for subtractive hybridization. Clones of the subtractive library were sequenced and the obtained sequences were identified by gen bank assignment. Two clones were found to contain sequences which could be assigned to the osmolarity sensor protein envZ, showing homologies of 61 and 65%. EnvZ is an inner membrane protein in bacteria, important for osmosensing and required for porine gene regulation. It undergoes autophosphorylation and subsequently phosphorylates OmpR, which in turn binds to the porin (outer membrane protein) promoters to regulate the expression of OmpF and OmpC, major outer membrane porines. This is the first report of an osmosensing mechanism in the mammalian system, which was described in bacteria only. Furthermore, we are assigning a tentative role for taurine in the osmoregulatory process by modifying the expression of the osmoregulatory sensor protein ENVZ.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1438-2199
    Keywords: Amino acids ; Half-specific PCR ; Thioredoxin ; Gene isolation ; Streptomyces aureofaciens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The method of two-stage half-specific amplification was described and successfully used in the isolation of the protein-coding part of the thioredoxin gene from Streptomyces aureofaciens BMK. The efficiency of a new PCR modification for the specific amplification of the target DNA fragments (genes) with unknown sequences is compared with the used half-specific PCR. The determined target sequence demonstrates the highest homology with the thioredoxin genes from Corynebacterium nephridii C-1 and Anabaena 7119.
    Type of Medium: Electronic Resource
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