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研究生:黃開成
研究生(外文):Huang, KaiCheng
論文名稱:植物乳桿菌穿梭表現載體之構築
論文名稱(外文):Construction of shuttle expression vector for Lactobacillus plantarum
指導教授:歐柏榮
指導教授(外文):Ou, BorRung
口試委員:歐柏榮
口試委員(外文):Ou, BorRung
口試日期:2011-06-15
學位類別:碩士
校院名稱:東海大學
系所名稱:畜產與生物科技學系
學門:農業科學學門
學類:畜牧學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:中文
論文頁數:106
中文關鍵詞:乳酸桿菌植物乳桿菌穿梭表現載體
外文關鍵詞:LactobacillusLactobacillus plantarumshuttle expression vector
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Lactobacillus genus is the largest group of lactic acid bacteria and plays an important role on animal health and food fermentation. Several researches used Lactobacillus to express exogenous proteins for scientific and industrial application. To develop the protein expression system for the manipulation of genetic engineering in Lactobacillus, a shuttle expression vector based on the region of replication origin derived from endogenous plasmid of Lactobacillus was constructed. This shuttle expression vector was able to replicate in both Escherichia coli and Lactobacillus to express exogenous proteins. The recombinant shuttle expression vector contains pGLO, an Escherichia coli cloning vector with GFP reporter gene, as the backbone, the p256 minimal replicon derived from endogenous plasmid in Lactobacillus plantarum ATCC 14917, and the erythromycin resistance gene (Ermr gene) amplified from pAMβ1 which is a Gram-positive wide-host range plasmid originated from Streptococcus faecalis DS5. The promoter sequence upstream of the GFP gene was replaced by slpA promoter amplified from Lactobacillus acidophilus ATCC 4356 genomic DNA to complete the construction of the shuttle expression vector, pSLP256gfp. The results showed Lactobacillus plantarum ASLA-1 transformed with pSLP256gfp successfully expressed GFP fluorescence under erythromycin selection and the existence of pSLP256gfp in the transformed host was confirmed by PCR analysis. In conclusion, the results from this study indicated that this shuttle expression vector can be further used to express exogenous proteins in Lactobacillus plantarum.
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壹、摘要-----------------------------------------------------------------------1

貳、序言-----------------------------------------------------------------------2

參、文獻回顧-------------------------------------------------------------------3

肆、材料與方法----------------------------------------------------------------28

伍、結果----------------------------------------------------------------------48

陸、討論----------------------------------------------------------------------82

柒、結論----------------------------------------------------------------------94

捌、參考文獻------------------------------------------------------------------95

玖、英文摘要-----------------------------------------------------------------105

拾、作者小傳-----------------------------------------------------------------106

捌、參考文獻

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