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Molecular cloning and nucleotide sequences of cDNAs specific for rat liver ribosomal proteins S17 and L30
Authors:O Nakanishi  M Oyanagi  Y Kuwano  T Tanaka  T Nakayama  H Mitsui  Y Nabeshima  K Ogata
Affiliation:1. Department of Biology Faculty of Science Niigata University Niigata 950-21, Japan Tel. (0252)-62-6174;1. Department of Biochemistry Niigata University School of Medicine Niigata 951, Japan Tel. (0252)-23-6161;2. Department of Biochemistry Yamagata University School of Medicine Yamagata 990-23, Japan Tel. (0236)-33-ll22;3. Department of Biochemistry Miyazaki Medical College Miyazaki 889-16, Japan Tel. (09858)-5-1510
Abstract:cDNA clones coding for rat liver ribosomal proteins S17 and L30 have been isolated by positive hybridization-translation assay from a cDNA library prepared from 8-9S poly(A)+RNA from free polysomes of regenerating rat liver. The cDNA clone specific for S17 protein (pRS17-2) has a 466-bp insert with the poly(A) tail. The complete amino acid (aa) sequence of S17 protein was deduced from the nucleotide sequence of the cDNA. S17 protein consists of 134 aa residues with an Mr of 15 377. The N-terminal aa sequence of S17 protein determined by automatic Edman degradation is consistent with the sequence data. The aa sequence of S17 shows strong homology (76.9%) to that of yeast ribosomal protein 51 [Teem and Rosbash, Proc. Natl. Acad. Sci. USA 80 (1983) 4403-4407] in the two-thirds N-terminal region. The cDNA clone specific for L30 protein (pRL30) has a 394-bp insert. The aa sequence of L30 protein was deduced from the nucleotide sequence of the cDNA. The protein consists of 114 aa residues with an Mr of 12 652. When compared with the N-terminal aa sequence of rat liver L30 protein [Wool, Annu. Rev. Biochem. 48 (1979) 719-754], pRL30 was found not to contain the initiation codon and 5'-noncoding region. The cDNA showed twelve silent changes in the coding region, one point mutation and one base deletion in the 3'-noncoding region, compared with mouse genomic DNA for L30 protein [Wiedemann and Perry, Mol. Cell Biol. 4 (1984) 2518-2528].
Keywords:Hybridization-translation assay  recombinant plasmid  Maxam-Gilbert technique  automatic Edman degradation  aa  amino aeid(s)  bp  base pair(s)  cDNA  DNA complementary to mRNA  ds  double stranded  nt  nucleotide(s)  PA  polyacrylamide  Pu  purine  resistant  rpe-mRNA  mRNA enriched for ribosomal protein-coding sequences  Tc  tetracycline
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