cDNA, genomic cloning and sequence analysis of ribosomal protein S4X gene (RPS4X) from Ailuropoda melanoleuca
Ribosomal protein S4X (RPS4X) is one of the 40S ribosomal proteins encoded by the RPS4X gene. The cDNA and the genomic sequence of RPS4X were cloned successfully from giant panda (Ailuropoda melanoleuca) using reverse transcriptase-polymerase chain reaction (RT-PCR) and touchdown-PCR technology respectively, followed by sequencing and analyzing. The cDNA of the RPS4X cloned is 812 bp in size, containing an open reading frame (ORF) of 792 bp encoding 263 amino acids with deduced molecular weight of 29.6 kD and isoelectric point (PI) of 10.16. The length of the genomic sequence is 5074 bp, which was found to possess 7 exons and 6 introns. There are 1 N-glycosylation site, 3 protein kinase C phosphorylation sites, 4 casein kinase II phosphorylation sites, 2 tyrosine kinase phosphorylation sites and 4 N-myristoylation sites in the RPS4X of giant panda by topology prediction. Alignment analysis indicated that the coding sequence and the deduced amino acid sequence of RPS4X gene shows very high homology with other reported mammalian species. The information on cDNA and the genomic sequence from giant panda RPS4X will provide basic information for further research of mammals RPS4X gene, which would be beneficial in the study of ribosomal biogenesis and would allow the elucidation of structure, organization and regulation of genes encoding ribosomal proteins in eukaryote.
Key words: RPS4X gene, giant panda, genomic cloning, sequence analysis.