合肥工业大学学报（自然科学版）
閤肥工業大學學報（自然科學版）
합비공업대학학보（자연과학판）
Journal of Hefei University of Technology (Natural Science)
2015年
9期
1277-1280
,共4页
钝齿棒杆菌%乳酸脱氢酶%基因敲除%载体构建
鈍齒棒桿菌%乳痠脫氫酶%基因敲除%載體構建
둔치봉간균%유산탈경매%기인고제%재체구건
Corynebacterium crenatum%lactate dehydrogenase%gene knock-out%vector construction
乳酸脱氢酶催化丙酮酸生成乳酸是钝齿棒杆菌厌氧发酵产乳酸的关键酶。文章以构建乳酸脱氢酶基因敲除载体为目的，通过在乳酸脱氢酶基因片段中的EcoRV酶切位点插入卡那霉素抗性基因的方法构建钝齿棒杆菌乳酸脱氢酶基因敲除载体。结果显示，成功扩增并克隆到乳酸脱氢酶基因与卡那霉素抗性基因，质粒聚合酶链式反应（polymerase chain reaction ，PCR）与酶切结果显示，卡那霉素抗性基因准确插入到乳酸脱氢酶基因中的EcoRV酶切位点，钝齿棒杆菌乳酸脱氢酶基因敲除载体构建成功。
乳痠脫氫酶催化丙酮痠生成乳痠是鈍齒棒桿菌厭氧髮酵產乳痠的關鍵酶。文章以構建乳痠脫氫酶基因敲除載體為目的，通過在乳痠脫氫酶基因片段中的EcoRV酶切位點插入卡那黴素抗性基因的方法構建鈍齒棒桿菌乳痠脫氫酶基因敲除載體。結果顯示，成功擴增併剋隆到乳痠脫氫酶基因與卡那黴素抗性基因，質粒聚閤酶鏈式反應（polymerase chain reaction ，PCR）與酶切結果顯示，卡那黴素抗性基因準確插入到乳痠脫氫酶基因中的EcoRV酶切位點，鈍齒棒桿菌乳痠脫氫酶基因敲除載體構建成功。
유산탈경매최화병동산생성유산시둔치봉간균염양발효산유산적관건매。문장이구건유산탈경매기인고제재체위목적，통과재유산탈경매기인편단중적EcoRV매절위점삽입잡나매소항성기인적방법구건둔치봉간균유산탈경매기인고제재체。결과현시，성공확증병극륭도유산탈경매기인여잡나매소항성기인，질립취합매련식반응（polymerase chain reaction ，PCR）여매절결과현시，잡나매소항성기인준학삽입도유산탈경매기인중적EcoRV매절위점，둔치봉간균유산탈경매기인고제재체구건성공。
When producing lactic acid by Corynebacterium crenatum under anaerobic condition ,the lac‐tate dehydrogenase is the key enzyme which can catalyze the formation of lactic acid from pyruvate .In this paper ,the ldhA gene knock‐out vector was constructed by inserting a km gene into the EcoRV re‐striction site of ldhA .The results indicated that the ldhA and km genes were cloned successfully ,the positive recombinant plasmid was identified by polymerase chain reaction(PCR) and enzyme digestion and the results certified that the km gene was accurately inserted into the EcoRV restriction site of ldhA and the ldhA gene knock‐out vector was successfully constructed .
