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从狐狸睾丸cDNA文库中克隆、测序并鉴定乳酸脱氢酶C4

Cloning, sequencing, and characterization of LDH-C4 from a fox testis cDNA library.

作者信息

Bradley M P, Geelan A, Leitch V, Goldberg E

机构信息

Perth Zoological Gardens, South Perth, Western Australia, Australia.

出版信息

Mol Reprod Dev. 1996 Aug;44(4):452-9. doi: 10.1002/(SICI)1098-2795(199608)44:4<452::AID-MRD4>3.0.CO;2-K.

Abstract

A full-length cDNA encoding the sperm-specific enzyme lactate dehydrogenase-C4 was isolated from a fox testis cDNA expression library and sequenced. The deduced translated protein sequence was shown to be 86% identical to that of human LDH-C4. In the fox testis, mRNA encoding LDH-C4 was first detected in pachytene spermatocytes. The LDH-C4 protein monomer was identified in Western blots of sperm membrane extracts as having a molecular weight of approximately 35,000, consistent with the monomeric size of this subunit previously identified in sperm from other species. The LDH-C4 protein is localized on the sperm plasma membrane overlying the principal piece of the tail. Based on the available sequence data, we were able to identify an epitope within the N-terminal region of the LDH-C4 amino-acid sequence which when administered to female foxes is antigenic and produces antibodies capable of recognizing the native protein.

摘要

从狐狸睾丸cDNA表达文库中分离出编码精子特异性酶乳酸脱氢酶-C4的全长cDNA并进行测序。推导的翻译后蛋白质序列与人LDH-C4的序列有86%的同一性。在狐狸睾丸中,编码LDH-C4的mRNA首先在粗线期精母细胞中被检测到。在精子膜提取物的蛋白质印迹中鉴定出LDH-C4蛋白单体,其分子量约为35000,与先前在其他物种精子中鉴定的该亚基的单体大小一致。LDH-C4蛋白定位于覆盖尾部主段的精子质膜上。基于现有的序列数据,我们能够在LDH-C4氨基酸序列的N端区域鉴定出一个表位,当将其给予雌性狐狸时具有抗原性,并产生能够识别天然蛋白的抗体。

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