臺灣博碩士論文加值系統

到目前為止，我們只知 DNase 型大腸桿菌素進入細菌時需要 BtuB、OmpF、與 Tol 等蛋白質的協助，但是根據這些蛋白質所提出的胞膜移位模式，對於大腸桿菌素如何與免疫蛋白質解離，以及進入內膜的機制，解釋仍不夠詳盡。為了研究大腸桿菌素 E7 進入細菌的過程中是否有其他分子參與，以及這些分子的功能，本實驗利用轉位子 Tn5 送入大腸桿菌 K12 菌株，進行轉位突變，從中篩選出 56 株對大腸桿菌素 E7 有抗性的菌株，並且分析了其中 36 株的突變基因，以了解這些突變株產生抗性的機制，並探討這些突變基因對大腸桿菌素胞膜移位過程的影響。其他位於胞膜間區的蛋白質也可能與大腸桿菌作用，我們選用 ompT、pcr 等蛋白酶突變株，以及 dnaJ chaperonin 與 malE maltose transpoter 的突變株來測試對大腸桿菌素 E7 的抗性，結果顯示這些突變株與對大腸桿菌素 E7 的抗性無關，因此大腸桿菌素 E7 進入細菌時並不需要這些蛋白質協助，然而對於大腸桿菌素 E7 向外運送時是否有影響仍需進一步證明。另外，根據一部份突變株的實驗結果得知：大腸桿菌 K12 菌株與 O157 菌株皆能利用 OmpC 與 OmpF 進入細菌，而二者表現量會互為消長，目前已知大腸桿菌素 N 使用 OmpC 進入細菌的效率較差，因此我們推測某些突變株抵抗大腸桿菌素 E7 的機制可能是藉由增加 OmpC 的表現，並減少細菌外膜 OmpF 的數量，使大腸桿菌素 E7 不易進入細菌。

It is well known that BtuB, OmpF, and Tol proteins are required for the import of DNase-type colicins. However, the import mechanism established by these proteins cannot elucidate ow the colicins are dissociated with the immune protein and pass through the inner membrane of the bacteria. It is possible that some new genes may be mutagenesis method was used. Fifty-six colicin-resistant mutants were obtained and then thirty-six mutated genes of these mutants were analyzed to study the roles of these genes played in the mechanism of colicin-resistance and their influences on clilcin-translocation. Other periplasmic proteins, such as protease, chaperon, and transporters, may alsi be involved in the translocation of colicins. To investigate this possibility, the E. coli strains with ompT, per, dnaJ, or malE mutation were tested respectively for the colicin-resistance. The results showed that these four genes were not involved in the colicin E7-resistance. Furthermore, the analysis of ompC mutants of E. coli strains K12 and O157 revealed that not only OmpF but also OmpC might help colicin E7 to translocate the membrane. Since the entering rate of colicin N via OmpC is lower than OmpF, it is proposed that the increase of OmpC and reduction of OmpF may be a benefit to the host for colicin-resistance.

目錄...................1
英文摘要...............3
中文摘要...............4
背景介紹...............6
材料與方法............20
結果..................32
討論..................38
圖表..................50
參考文獻..............58

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夏國強. (2000). 陽明大學生化所碩士論文：大腸桿菌素與胞膜間區蛋白質交互作用之研究.
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