在大腸桿菌中，離胺酸的合成需要經過九個步驟。天門冬胺酸磷酸化酵素III是負責離胺酸合成路徑中的第一步：將天門冬胺酸予以磷酸化。在此論文中，我們利用噬菌體的重組去氧核糖核酸系統成功剔除天門冬胺酸磷酸化酵素III的基因lysC，接著我們利用基因晶片的技術比較lysC 剔除與野生種大腸桿菌 K-12 W3110 之所有的基因表現變化的差異。結果發現，有許多基因有明顯的表現差異，包括與離胺酸合成有關的asd、 dapB 與dapD。 另外， 與天門冬胺酸合成有關的基因ppc 及gdhA也有明顯的上升表現。 在 lysC 剔除的大腸桿菌中， 負責離胺酸運輸的蛋白其基因lysP也有顯著的上升.基於上述的結果，我們推論lysC 剔除的大腸桿菌是處在離胺酸缺乏的情況。之後我們為了進一步了解lysC基因對離胺酸合成途徑上其他基因的影響，同時了解它與大腸桿菌中其他生化合成系統的影響，我們設計了一套時間系列的生物晶片實驗來觀察野生型與lysC剔除的大腸桿菌在面對離胺酸拮抗劑S-(2-aminoethyl) cysteine (AEC) 的反應。結果顯示AEC造成許多影響，包括離胺酸的合成、核苷酸的合成、硫基的代謝、運輸系統與緊迫蛋白的基因表現。AEC的處理造成了離胺酸過剩的表現型，與合成離胺酸有關的基因如asd、 dapA、 dapB、 dapD 及lysA之表現都明顯的受到壓抑，同時與天門冬胺酸合成有關的基因aspC、 ppc、 gdhA也都被發現減少其表現量。本論文利用生物晶片的方式有助於釐清了lysC基因在大腸桿菌中扮演的系統角色，並進一步幫助了解大腸桿菌在合成離胺酸途徑上的基因調控。

In Escherichia coli, the biosynthesis of lysine is generated from aspartate, and the process contains nine steps. Aspartokinase III encoded by lysC, catalyzes the first step of lysine biosynthesis. In this thesis, we generated a lysC knockout strain E. coli K-12 W3110 using汹财 red recombination system. And we inspected the global gene expression profiles between lysC knockout and wild type E. coli. Several significant differential gene expressions were observed, including the up-regulation of lysine biosynthetic pathway genes like asd, dapB and dapE. Genes related to aspartate and glutamate biosynthetic processes like ppc and gdhA were up-regulated as well. These results suggested that the lysC knockout exhibited a lysine starvation phenotype. A time series microarray analysis was carried out following the lysC knockout profiling experiment. S-(2-aminoethyl) cysteine (AEC) was added to the medium and the gene expression profiles following four time points were monitored. The results of AEC treatment time series expression analysis showed that the lysC-induced lysine starvation phenotype was neutralized by AEC. AEC affected several clusters of gene expressions, including lysine biosynthetic process, nucleotide biosynthetic process, sulfate metabolic process, transport system and stress response proteins. AEC treated lysC knockout exhibited a lysine excess phenotype by repressing the lysine biosynthetic process genes like asd, dapA, dapB, dapD and lysA. Genes related to aspartate biosynthesis like aspC and ppc and gdhA were also found down-regulated after AEC treatment. The results of this thesis help clarifying the systematic regulation of lysine biosynthesis in E. coli.

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