title: cDNA, genomic sequence cloning  and overexpression of giant panda (Ailuropoda  melanoleuca) mitochondrial ATP synthase ATP5G1.
abstract: The ATP5G1 gene is one of the three genes that encode mitochondrial ATP synthase  subunit c of the proton channel. We cloned the cDNA and determined the genomic  sequence of the ATP5G1 gene from the giant panda (Ailuropoda  melanoleuca) using RT-PCR technology and touchdown-PCR, respectively. The  cloned cDNA fragment contains an open reading frame of 411 bp encoding 136  amino acids; the length of the genomic sequence is of 1838 bp, containing three  exons and two introns. Alignment analysis revealed that the nucleotide sequence  and the deduced protein sequence are highly conserved compared to Homo  sapiens, Mus musculus, Rattus norvegicus, Bos taurus,  and Sus scrofa. The homologies for nucleotide sequences of the giant  panda ATP5G1 to those of these species are 93.92, 92.21, 92.46, 93.67, and  92.46%, respectively, and the homologies for amino acid sequences are 90.44,  95.59, 93.38, 94.12, and 91.91%, respectively. Topology prediction showed that  there is one protein kinase C phosphorylation site, one casein kinase II  phosphorylation site, five N-myristoylation sites, and one ATP synthase c  subunit signature in the ATP5G1 protein of the giant panda. The cDNA of ATP5G1 was transfected into Escherichia coli, and the ATP5G1 fused with the  N-terminally GST-tagged protein gave rise to accumulation of an expected 40-kDa  polypeptide, which had the characteristics of the predicted protein.
authors.affiliation: Key Laboratory of Southwest China Wildlife Resources Conservation, Ministry of Education, College of Life Science, China West Normal University, P.R. China. hwr168@yahoo.com.cn
authors.forename: W-R
authors.initials: WR
authors.lastname: Hou
authors.forename: Y-L
authors.initials: YL
authors.lastname: Hou
authors.forename: X
authors.initials: X
authors.lastname: Ding
authors.forename: T
authors.initials: T
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D016678:Genome / Q000235:genetics*; D007526:Isoelectric Point; D025261:Mitochondrial Proton-Translocating ATPases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity; D001503:Ursidae / Q000235:genetics*
chemical_list: D018076:DNA, Complementary; D025261:Mitochondrial Proton-Translocating ATPases