cDNA, Genomic Sequence Cloning and Sequence Analysis of Heat Shock Protein Beta-1 Gene (HSPB1) from the Giant Panda (Ailuropoda melanoleuca)

摘要

The HSPB1'office full name is heat shock protein beta-1, which is well-defined as a stress-inducible protein. It is a significant and interesting work to study it . The cDNA and the genomic sequence of HSPB1 was cloned successfully from the Giant Panda (Ailuropoda melanoleuca) using RT-PCR technology and touchdown-PCR, respectively, Both sequences were sequenced and analyzed preliminarily. The cDNA of the HSPB1 cloned from Giant Panda is 633bp in size, containing an open reading frame (ORF) of 624bp encoding 207 amino acids. The length of the genomic sequence is 1193 bp, which was found to possess three exons and two introns. Alignment analysis indicates that the nucleotide sequence of the coding sequence shows a high homology to those of Homo sapiens, Mus musculus, Bos Taurus, Canis lupus familiaris, Rattus norvegicus and Equus caballus are 89%, 85%, 88%,93%,83% and 91%, respectively . The homologies for deduced amino acid sequence are 89%, 90%,90%, 92%,88% and 92%, respectively. Topology prediction shows there are two N-myristoylation sites, four Protein kinase C phosphorylation sites, one Tyrosine kinase phosphorylation site, one cAMP- and cGMP-dependent protein kinase phosphorylation site and a ATP/GTP-binding site motif A (P-loop) in the HSPB1 protein of the Giant Panda. Futher analysis indicates that the expression sequence of HSPB1and the protein encoded are highly homologous to those of human and other mammals reported.