CDNA and genomic sequence clonging and sequence analysis of ribosomal protein S5 gene(rpS5) from giant panda and its overexpression

Abstract

RPS5 is a component of the 40S small ribosomal subunit encoded by rpS5 gene, which is specific to eukaryotes. The cDNA and the genomic sequence of rpS5 were cloned successfully from giant panda using RT-PCR technology and Touchdown-PCR, respectively. Both sequences were analyzed preliminarily and the cDNA of the rpS5 gene was overexpressed in E. coli BL21. The cDNA of rpS5 cloned from giant panda contain open reading frame of 615 bp encoding 204 amino acids. The nucleotide sequence of the coding sequence shows a high homology to some mammals as determined by Blast analyzing, and the same with the amino acid sequence. The genomic sequence is 4331 bp in length, with five exons and four introns. Primary structure analysis revealed that the molecular weight of the putative RPS5 protein is 22.87 KD with a theoretical pI 10.30. Topology prediction showed there are five different patterns of functional sites in the RPS5 protein of giant panda. The rpS5 can be really expressed in E. coli and its protein fused with the N-terminally GST -tagged protein gave rise to the accumulation of an expected 29.5 KD polypeptide. The expression product obtained could be used for purification and study of its function further.