cDNA, genomic sequence cloning and overexpression of giant panda (Ailuropoda melanoleuca) mitochondrial ATP synthase ATP5G1

Abstract

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 ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 11 (3): 3164-3174 (2012) ATP5G1 of the giant panda 3165 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.

The correlated expression of immune and energy metabolism related genes in the response to Salmonella enterica serovar Enteritidis inoculation in chicken

“…ATP5G1, a key component of complex V of the oxidative phosphorylation chain, encodes a subunit of mitochondrial ATP synthase and catalyzes ATP synthesis [ 29 ] and is involved in the biological process of oxidative phosphorylation and might be associated with oxidative stress [ 30 ]. The expression of ATP5G1 and ATP5G3 is actively regulated in response to various physiological stimuli, and maintain the basal levels of the c-Fo subunit [ 31 ]. However, little information is available on the regulation of S .…”

Molecular cloning, overexpression, purification, and sequence analysis of the giant panda (Ailuropoda melanoleuca) ferritin light polypeptide

“…Over recent years, conservation measures have been put in place and scientists have realized tremendous success in areas such as breeding, ecology, and propagation, among others. Meanwhile, giant panda research and conservation has been increasingly focusing on the level of gene functionality, which is currently drawing much attention in this field (Allerson et al, 1999;Liao et al, 2003;Hou et al, 2012;Hou et al, 2013;Wang et al, 2015;Wang et al, 2016). In this study, we designed primers based on the available FTL gene sequences of several mammalian species, including Homo sapiens, Cavia porcellus, Equus caballus, and Felis catus, and used reverse transcriptase-polymerase chain reaction (RT-PCR) to amplify FTL complementary DNA (cDNA) from total RNA isolated from giant panda skeletal muscle.…”