- 學位論文
大白鼠儲精囊蛋白RSVS III基因及其功能研究
Genetic and Functional Study of the Rat Seminal Vesicle Secretion Protein III, RSVS III
摘要
大白鼠儲精囊分泌主要蛋白依分子量依序為RSVS I~V，本實驗室曾將RSVS III以質譜儀定序的結果顯示此蛋白為大白鼠RpSv-1 mRNA所轉譯的產物。 由免疫組織化學法的檢視，顯示RSVS III是由儲精囊表皮細胞分泌累積在腔道。 RSVS III 蛋白可以和RSVS I及 RSVS II受到transglutaminase催化而形成高分子的共價聚合物，顯示RSVS III是交配栓的組成蛋白之一。 RpSv-1(RSVS III?基因的結構包含啟動子區序列 1771個鹼基對，3個長度分別為92、1482及327個鹼基對的exon和2個長度分別為230及89個鹼基對的intron。其中，exon1轉譯為signal peptide，exon2轉譯為完整的蛋白質序列，而exon3是3’-untranslated region，此基因結構特色符合「快速演化的轉麩胺醯胺酸基質(Rapid Evolving Substrate of Transglutaminase)」(REST)基因家族。 RSVS IIIㄟ穧]位於大白鼠染色體3q42，在此染色體區域中，發現另一個和RpSv-1(RSVS III?基因序列99.9%相似的RSVS IIIβ基因，以北方墨點法及RT-PCR確認這兩個基因都專一地表現在儲精囊中，其mRNA表現皆受到雄性素的刺激，然而兩基因的表現量不同。儲精囊表現的REST基因在人類染色體20q13、小白鼠染色體2H3及大白鼠染色體3q42，其中後二者有相似的基因體排列，我以這些基因的序列分析及演化樹的建構探討其分子演化的關係。
並列摘要
Rat seminal vesicle secretion protein contains five well-defined major components designated RSVS I-V in decreasing order of molecular weight. RSVS I-III were covalent cross-linked to form high molecular complexes by transglutaminase. RSVS III was confirmed to be derived from RpSV-1 mRNA. RSVS III was immunolocalized in the luminal epithelium of seminal vesicle and the copulatory plug. The genomic structure of RpSv-1(RSVS III?was analyzed to establish a 5’-flanking region of 1771 bp, three exons of 92, 1482 and 327 bp, and two introns of 230 and 89 bp. The transcription unit is organized with the first exon encoding a signal peptide, and the second exon encoding a protein in its entirety, whereas the third encompasses a 3’-untranslated region, that shares common features with the arrangement of transcription units of the rapid evolving substrate of tansglutaminase (REST) gene family. The RSVS III?gene was mapped to chromosome 3q42 locus. We found another paralog, RSVS IIIβ gene, which shares 99.9% nucleotide sequence identity to RSVS III?in the same locus. Northern blotting and RT-PCR were performed to demonstrate the exclusive expression of this duplicated pair in seminal vesicle with a ratio of RSVS III?mRNA to RSVS IIIβ mRNA ≒ 2.0. The mRNA expression of RSVS III? RSVS IIIβ were stimulated by testosterone. The genes in human chromosome 20q13, mice chromosome 2H3 and rat chromosome 3q42 were identified to have the arrangement of their transcription units similar to a REST gene. The molecular evolution of these REST genes is discussed.