Cloning and sequence analysis of ribosomal protein L8 gene (RPL8) from the (Ailuropoda melanoleuca)

Abstract

Ribosomal protein L8 is a component of the 60S ribosomal subunit, encoded by the RPL8 gene in eucaryotic cell. The protein belongs to the L2P family of ribosomal proteins. To explore the structure characteristic of RPL8 gene of the Giant Panda, Ailuropoda melanoleuc), we have designed primers based on the known nucleotide sequence of RPL8 genes to clone the cDNA and genomic sequences of RPL8 gene of A. melanoleuca by RT-PCR and PCR strategy respectively, and then the cloned cDNA and genomic sequences were sequenced and analyzed preliminarily. The results indicated that the length of cDNA fragment of the RPL8 from the Giant Panda was 814 bp in size, containing an open reading frame (ORF) of 774 bp, encoding 257 amino acids. The length of the genomic sequence was 2,370 bp, with five exons and four introns. The coding sequence showed a high degree of homology to those of Homo sapiens, Bos taurus, Canis lupus, Mus musculus and Rattus norvegicus by 92.12, 90.11, 95.99, 86.46, and 88.50%, respectively. The homologies for the deduced amino acid sequences reached high up to 100%. Primary structure analysis revealed that the molecular weight of the putative RPL8 protein was 28.02 kD, with a theoretical pI of 11.65. Based on topology prediction, there were one cAMP- and cGMP-dependent protein kinase phosphorylation site, four protein kinase C phosphorylation site, two casein kinase II phosphorylation site, seven N-myristoylation sites, two Amidation site and one Ribosomal protein L2 signature in the RPL8 protein of the Giant Panda.