Int J Biol Sci 2010; 6(4):316-326. doi:10.7150/ijbs.6.316
Cloning, characterization, and expression analysis of goat (Capra hircus) phospholipid hydroperoxide glutathione peroxidase (PHGPx)
1. College of Animal Science and Veterinary Medicines, Shanxi Agricultural University, Taigu, Shanxi 030801, China
2. College of Animal Science and Veterinary Medicine; Taiyuan Shanxi 030024, China
3. Licheng sheep Breeding Base of Shanxi Agricultural University, Licheng Shanxi 047600, China
SHI Lg, XUN Wj, YUE Wb, ZHANG Cx, REN Ys, WANG Q, WU Xy, SHI L, YANG Rj, LEI Fl. Cloning, characterization, and expression analysis of goat (Capra hircus) phospholipid hydroperoxide glutathione peroxidase (PHGPx). Int J Biol Sci 2010; 6(4):316-326. doi:10.7150/ijbs.6.316. Available from http://www.ijbs.com/v06p0316.htm
Phospholipid hydroperoxide glutathione peroxidase (PHGPx), as a ubiquitous antioxidant enzyme in the glutathione peroxidases (GPx) family, plays multiple roles in organisms. However, there is very little information on PHGPx in goats (Capra hircus). In this study, a full-length cDNA was cloned and characterized from Taihang black goat testes. The 844 bp cDNA contains an open reading frame (ORF) of 597 bp. The goat PHGPx nucleotide sequence contains a selenocysteine (sec) codon TGA244-246, two potential start codons ATG20-22 and ATG108-110, a polyadenylation signal AATAAA813-818 and selenocysteine insertion sequence (SECIS) motif AUGA688-691, UGA729-731 and AAA703-705. As a selenoprotein, the active-site motifs and GPx family signature motifs LAFPCNQF101-108 and WNFEK165-170 were also found. The order of PHGPx mRNA expression levels was: testes >> heart > brain > epididymis > kidney > liver > lung > spleen > muscle. Real-time PCR and immunohistochemistry results revealed similar expression differences in different age testes, with high expression levels during adolescence. Immunofluorescence results suggested that PHGPx mainly expressed in Leydig cells and spermatids in mature goat testes.
Keywords: goat, phospholipid hydroperoxide glutathione peroxidase, real-time PCR, expression, immunohistochemistry.