小白鼠貯精囊分泌液中之蛋白脢抑制因數之生物特性研究及其表現

Abstract

本實驗室自小白鼠貯精囊分泌液中純化出一種kazal-type trypsin inhibitor (TI)，經氨基酸定序結果與1987年Mills等人所找到的MP12 cDNA所推衍之蛋白質相同。MP12在胰臟中是持續表現，但在雄性生殖器官卻須雄性激素誘導才會表現，該蛋白質的功能並不十分清楚。 本篇論文主要在探討Tl在小老鼠器官分佈情形，受雄性素調控的現象，以及在不同年齡TI量的變化，並利用遺傳工程的方法，將用PCR放大之包含成熟TI的DNA片段建構於pGEX載體中，於E. coli 中表現，然後用affinity collumn回收fusion-TI，做活性分析。

A Kaza-type trypsin inhibitor(TI)was purified from mouse seminal vesicle. The amino acid sequence of this protein is identical to the protein sequence deduced from MP12 cDNA sequence. MP12 mRNA requires androgen for its expression in male sex accessory tissues but expressed constitutively in pancreas. The physiological role of this protein is still unclear. In the present study, the tissue distribution of TI, androgen-dependentin seminal vesicle and the level of TI in different ages of mouse were investigated. By using genetic engineering, the DNA fragment containing mature TI was amplified through Polymerase Chain Reaction(PCR), constructed into pGEX2 vector, and expressed in E. coli. The fusion protein was purified by affinity beads and trypsin inhibition activity was tested.