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Cloning, characterization, and expression analysis of goat (Capra hircus) phospholipid hydroperoxide glutathione peroxidase (PHGPx)

机译:山羊(Capra hircus)磷脂氢过氧化物谷胱甘肽过氧化物酶(PHGPx)的克隆,鉴定和表达分析

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摘要

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.
机译:磷脂氢过氧化物谷胱甘肽过氧化物酶(PHGPx),作为谷胱甘肽过氧化物酶(GPx)家族中普遍存在的抗氧化酶,在生物体中起着多种作用。但是,关于山羊(Capra hircus)中PHGPx的信息很少。在这项研究中,从太行黑山羊睾丸中克隆并鉴定了全长cDNA。 844 bp 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TGA244-246,两个潜在的起始密码子ATG20-22和ATG108-110,一个多腺苷酸化信号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表达水平的顺序为:睾丸>>心脏>脑>附睾>肾脏>肝脏>肺>脾脏>肌肉。实时PCR和免疫组织化学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。免疫荧光结果表明,PHGPx主要在成熟山羊睾丸的睾丸间质细胞和精子中表达。

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