本實驗室從國內健康成人腸道分離出一全新的乳酸菌種，Lactobacillus rhamnosus TCELL-1，未發現有內源性質體存在。本實驗是爲了構築一個能在此菌中表現的食品級選殖載體。
我們利用乳酸鏈球菌素免疫蛋白，nisI，作為篩選指標，並利用此菌的乳糖調控子的啟動子，pLac，作為nisI基因的啟動子，再利用乳酸菌廣域宿主複製源pAMβ1及pUC origin構築成一可在乳酸菌及大腸桿菌中複製的穿梭載體pNI。在此一穿梭載體中還接有一段nucT報導基因，作為篩選指標。
食品級選殖載體pNI經由電轉型方式送入Lb. rhamnosus TCELL-1內後，我們利用南方轉漬雜交法證實質體pNI確實存在於此菌內。再利用乳糖誘導表現ni I基因，測定此菌抵抗乳酸鏈球菌素的生長曲線，發現含有質體pNI的Lb. rhamnosus TCELL-1可以抵抗乳酸鏈球菌素達60 IU/ml，並利用plate diffusion assay測定此菌株與wild-type Lb. rhamnosus TCELL-1抗乳酸鏈球菌素之差別。

One new strain of Lactobacillus, identified as Lactobacillus rhamnosus TCELL-1, was isolated from the healthy adult rectum biopsies in our laboratory. A new food-grade shuttle cloning vector for Lb. rhamnosus TCELL-1 and E. coli was constructed using the nisin immunity gene nisI as a selection marker. The food-grade shuttle cloning vector, pNI, was constructed using the pAMβ1 replicon, the pUC origin, the nisI gene, the promoter Lac for nisI expression, and the nucT reporter gene. Electroporation into Lb. rhamnosus TCELL-1 was selected with the nucT reporter gene. Plasmid pNI was confirmed in Lb. rhamnosus TCELL-1 with southern hybridization. Lb. rhamnosus TCELL-1 carrying pNI was shown to be able to grow in medium containing a maximum of 60 IU nisin/ml. These results show that the food-grade expression system reported in this paper has potential for expression of foreign genes in Lb. rhamnosus TCELL-1 in order to construct improved starter bacteria for food applications.

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