在本研究中我們探討了新穎質體pSEI以何種方式銜接催乳素基因(PRL)，在送出廠商定序前如何初步確認有無做出預期的質體，PCR引子設計的考量、各種大腸桿菌菌株的使用目的以及菌株對質體表現的影響，包含蛋白的毒性測試、改善蛋白質表達方式，使目標蛋白可以在大腸桿菌系統中有更好的可溶性，增加後期純化的便利性。最後以His-tag做為主要的純化方法，搭配內含肽進行目標蛋白的分割，利用膠體電泳與螢光感測儀對目標蛋白的檢測數據做檢討。 The specific aims of this study are to investigate what way the new plasmid-pSEI uses to bridge Prolactin(PRL), and how to initially confirm whether the expected plasmid is made before senting the squencing. The research also shows the design considerstions of Polymerase Chain Reaction(PCR), the purpose of usage of every Escherichia coli (E.coli) and the influence that the strain towards the plasmid performance, including the test of toxic Protein, the improvement in the expression of the protein, making the target protein gives the good solubility in the E.coli system, and then increasing the convenience of the later purification. The experiment uses His-tag as a main way of the purification with the intein, dividing the target protein, then reflecting the data by testing the target protein with the Gel electrophoresis and the flurosensor.
