利用新穎質體優化人類催乳素蛋白於大腸桿菌的表達及其純化之研究

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Title:	利用新穎質體優化人類催乳素蛋白於大腸桿菌的表達及其純化之研究
Other Titles:	Optimizing the overexpression of human prolactin in E.coli with modified plasmid and purification.
Authors:	陳日賢 Chen-Rih-Sian
Contributors:	顧野松 Yesong Gu 化學工程與材料工程學系
Keywords:	大腸桿菌;催乳素;低溫表達;可溶性蛋白;His-tag純化;內含肽切割 E.coli;Prolactin;Low Temperature Expression;Soluble Protein;His-tag Purification
Date:	2019
Issue Date:	2019-12-16T02:19:00Z (UTC)
Abstract:	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.
Appears in Collections:	[Department of Chemical and Materials Engineering ] Theses and Dissertations

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