cDNA Cloning and Overexpression of Acidic Ribosomal Phosphoprotein P1 Gene (RPLP1) from the Giant Panda

Abstract

RPLP1 is one of acidic ribosomal phosphoproteins encoded by RPLP1 gene, which plays an important role in the elongation step of protein synthesis. The cDNA of RPLP1 was cloned successfully for the first time from the Giant Panda (Ailuropoda melanoleuca) using RT-PCR technology, which was also sequenced, analyzed preliminarily and expressed in E.coli. The cDNA fragment cloned is 449bp in size, containing an open reading frame of 344bp encoding 114 amino acids. Alignment analysis indicated that the nucleotide sequence and the deduced amino acid sequence are highly conserved to other five species studied, including Homo sapiens, Mus musculus, Rattus norvegicus, Bos Taurus and Sus scrofa. The homologies for nucleotide sequences of Giant Panda PPLP1 to that of these species are 92.4%, 89.8%, 89.0%, 91.3% and 87.5%, while the homologies for amino acid sequences are 96.5%, 94.7%, 95.6%, 96.5% and 88.6%. Topology prediction showed there are three Casein kinase II phosphorylation sites and two N-myristoylation sites in the RPLP1 protein of the Giant Panda (Ailuropoda melanoleuca). The RPLP1 gene was overexpressed in E. coli and the result indicated that RPLP1 fusion with the N-terminally His-tagged form gave rise to the accumulation of an expected 18kDa polypeptide, which was in accordance with the predicted protein and could also be used to purify the protein and study its function.

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

“…Previous studies on the Giant Panda have mainly concentrated on the macro level, such as breeding and propagation, ecology, genetic diversity, parentage, and so on [14][15][16]. Recently, researches on functional genes of giant panda are becoming a hot issue, especially in gene cloning and functional investigation such as ribosomal phosphoprotein P1 gene (RPLP1), RPS15, RPS19 and so on [17][18][19][20][21][22].…”

cDNA, Genomic Sequence Cloning, and Overexpression ofEIF1from the Giant Panda (Ailuropoda Melanoleuca) and the Black Bear (Ursus Thibetanus Mupinensis)

“…[13] Studies on the giant panda and the black bear have been concentrated on fields from breeding and propagation, ecology, et al [14][15][16][17][18][19] to molecular level. [20][21][22][23][24][25][26][27][28][29][30][31][32][33] EIF1 is therefore an essential and universally conserved translation factor with established functions and the study on the EIF1 already quite rich, but the mechanism of its action is still controversial, which has drawn our interest in the study, especially on the unreported EIF1of giant panda and black bear. We address this question by describing the structure of the protein and providing new data for genetic diversity, parentage, phylogenesis of these two species.…”

Comparative analysis and molecular characterization of genomic sequences and proteins of FABP4 and FABP5 from the giant panda (Ailuropoda melanoleuca)

“…It is at high risk of extinction due to human population expansion and conse-quent destruction of its habitat. At present, there is very little genetic information about the panda (Du et al, 2007;Hou et al, 2008a,b;Zhang et al, 2009), which is an essential tool for obtaining a detailed understanding of the biology of this organism. Recently, the giant panda's draft genome sequence, which has good coverage and completeness for genes with unique sequences, was generated and assembled ; however, some genes are still incomplete in this project.…”