Cloning, characterization, and expression analysis of goat (Capra hircus) phospholipid hydroperoxide glutathione peroxidase (PHGPx).
Cloning, characterization, and expression analysis of goat (Capra hircus) phospholipid hydroperoxide glutathione peroxidase (PHGPx).
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山羊 (Capra hircus) 磷脂氢过氧化物谷胱甘肽过氧化物酶 (PHGPx) 的克隆、表征和表达分析
DOI：
10.7150/ijbs.6.316
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发表时间：
2010-06-10
影响因子：
9.2
通讯作者：
Lei FL
中科院分区：
文献类型：
--
作者：
Shi LG;Xun WJ;Yue WB;Zhang CX;Ren YS;Wang Q;Wu XY;Shi L;Yang RJ;Lei FL
磷脂氢过氧化物谷胱甘肽过氧化物酶（PHGPx）是谷胱甘肽过氧化物酶（GPx）家族中普遍存在的一种抗氧化酶，在生物体中发挥着多种作用。然而，关于山羊（Capra hircus）中PHGPx的信息很少。本研究从太行黑山羊睾丸中克隆了一个全长cDNA，并对其进行了鉴定。全长844 bp的cDNA包含597 bp的开放阅读框（ORF）。山羊PHGPx核苷酸序列包含一个硒代半胱氨酸（sec）密码子TGA244-246、两个潜在启动密码子ATG20-22和ATG108-110、一个多聚腺苷化信号AATAAA813-818和硒代半胱氨酸插入序列（SECIS）基序AUGA688-691、UGA729-731和AAA703-705。作为硒蛋白，还发现了活性位点基序和GPx家族特征基序LAFPCNQF101-108和WNFEK165-170。PHGPx mRNA表达量的大小顺序为：睾丸>>心脏>脑>附睾>肾脏>肝脏>肺>脾脏>肌肉。Real-time PCR和免疫组化结果显示，不同年龄睾丸的表达差异相似，青春期表达水平较高。免疫荧光结果显示，PHGPx主要表达于成熟山羊睾丸间质细胞和精细胞中。
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.