...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular reproduction and development >Cloning, sequencing, and expression analysis of mouse glucosamine-6-phosphate deaminase (GNPDA/oscillin).
【24h】

Cloning, sequencing, and expression analysis of mouse glucosamine-6-phosphate deaminase (GNPDA/oscillin).

机译：小鼠6氨基葡萄糖氨基磷酸脱氨酶（GNPDA / oscillin）的克隆，测序和表达分析。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

It was reported previously that a hamster protein, called "oscillin," with a sequence related to that of an Escherichia coli glucosamine 6-phosphate deaminase (GNPDA) [glucosamine-6 phosphate isomerase] triggered Ca2+ oscillations in mammalian oocytes when introduced into their cytoplasm upon fertilization. Recently, it has been shown that GNPDA/oscillin is ubiquitously expressed in rat tissues and that a recombinant hamster GNPDA/oscillin protein does not exhibit oscillin activity when injected into oocytes. In mice, the nature and role of such a GNPDA/oscillin is not known, but another candidate protein, truncated kit tyrosine kinase (tr-kit), has been proposed as a sperm factor causing oocyte activation. In order to clarify this issue, we havecharacterized the mouse homologue of hamster and human GNPDA/oscillin, and examined its expression along with that of tr-kit. We report here the molecular cloning and sequencing of mouse GNPDA/oscillin, which shows over 96% identity with the hamster andhuman homologues. Using specific primers, we performed a reverse transcription-PCR analysis to determine the tissue distribution of mouse GNPDA/oscillin mRNA. Unlike tr-kit mRNA, which is expressed solely in mouse testis, GNPDA/oscillin mRNA is detectedin unfertilized oocytes and in all tissues examined including testis, heart, thymus, liver, ovary, uterus, kidney, spleen and lung. The protein itself was detected in all tissues examined by western blots. Indirect immunofluorescence studies, using an antibody raised against hamster GNPDA, demonstrate that GNPDA is lost with the acrosome reaction of mouse spermatozoa, is localized in the equatorial and neck regions of human spermatozoa and the post-acrosomal region of hamster spermatozoa. Our results thus indicate that mouse GNPDA/oscillin, the homologue of hamster oscillin, unlike tr-kit, does not exhibit some of the required characteristics expected from a putative sperm-derived oocyte-activating factor. The nucleotide sequence reported has been deposited with the GenBank database with the accession number AF160355.
机译：以前有报道称，一种仓鼠蛋白，称为“ oscillin”，其序列与大肠杆菌的氨基葡萄糖6-磷酸脱氨酶（GNPDA）[氨基葡萄糖-6磷酸异构酶]有关，在引入哺乳动物卵母细胞后会触发Ca2 +振荡。受精后。近来，已经表明GNPDA / oscillin在大鼠组织中普遍表达，并且重组仓鼠GNPDA / oscillin蛋白在注射入卵母细胞时不表现出oscillin活性。在小鼠中，这种GNPDA / oscillin的性质和作用尚不清楚，但已提出另一种候选蛋白截短的酪氨酸激酶（tr-kit）作为引起卵母细胞活化的精子因子。为了阐明这个问题，我们表征了仓鼠和人GNPDA / oscillin的小鼠同源物，并研究了其与tr-kit的表达。我们在这里报告了小鼠GNPDA / oscillin的分子克隆和测序，显示与仓鼠和人类同源物超过96％的同一性。使用特定的引物，我们进行了逆转录-PCR分析，以确定小鼠GNPDA / oscillin mRNA的组织分布。与tr-kit mRNA仅在小鼠睾丸中表达不同，GNPDA / oscillin mRNA在未受精的卵母细胞和所检查的所有组织（包括睾丸，心脏，胸腺，肝脏，卵巢，子宫，肾脏，脾脏和肺脏）中都可以检测到。通过蛋白质印迹在所有组织中检测到蛋白质本身。使用针对仓鼠GNPDA的抗体进行的间接免疫荧光研究表明，GNPDA随小鼠精子的顶体反应而丢失，位于人精子的赤道和脖子区域以及仓鼠的精子后区域。因此，我们的结果表明，与tr-kit不同，小鼠GNPDA / oscillin（仓鼠oscillin的同系物）未显示出推定的精子来源的卵母细胞激活因子所期望的某些所需特征。报告的核苷酸序列已保存在GenBank数据库中，登录号为AF160355。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号