Cloning and sequence analysis of ribosomal protein L23 gene from Ailuropoda melanoleuca

Ribosonal protein L23 is a component of the 60s large ribosomal subunit, and plays an important role in ribosome. To explore the structure characteristic of RPL23 gene of giant panda and investigate its homologies with other species reported, the cDNA and genomic sequence of RPL23 gene were cloned using RT-PCR and touchdown-PCR technology. The sequence data were analyzed by corresponding software. The results showed the length of cDNA fragment cloned was 457bp; containing an open reading frame of 423 bp. Deduced protein was composed of 144 amino acids with an estimated molecular weight of 14.86kD and pI of 11.19. The length of the genomic sequence was 5011 bp, containing five exons and four introns. The predicted protein has six different patterns of functional sites. The RPL23 gene can be readily expressed in E. coli. When it was fused with the N-terminally His-tagged protein, it gave rise to accumulation of an expected 20.5 kD polypeptide, in good agreement with the predicted molecular weight. Alignment analysis indicated that RPL23 was highly similarity with the reported species at the level of DNA and protein. Topology prediction showed that three different patterns were found in the RPL23 protein of giant panda. In this paper, the results provided scientific material for enriching and improving the mammals RPL23 gene database.