中国饲料
中國飼料
중국사료
CHINA FEED
2013年
23期
17-20,23
,共5页
抗菌肽%Dermadistinctin-Q1%原核表达%生物活性
抗菌肽%Dermadistinctin-Q1%原覈錶達%生物活性
항균태%Dermadistinctin-Q1%원핵표체%생물활성
antibacterial peptide%Dermadistinctin-Q1%prokaryotic expression%biological activity
试验对抗菌肽Dermadistinctin-Q1(DDQ1)进行原核表达,并对其生物活性进行检测。采用重叠区扩增基因拼接法,多步PCR获得抗菌肽DDQ1全基因,并使用原核表达载体pET-32a(+)构建工程菌。通过优化诱导表达时间和异丙基硫代半乳糖苷(IPTG)终浓度,实现融合蛋白表达条件最佳化。利用肠激酶从融合蛋白中分离抗菌肽DDQ1后,使用Ni离子亲和层析法进行纯化。进一步的生物活性检测表明,抗菌肽DDQ1具有广谱抗菌活性和一定的热、酸耐受性,是一类极具开发潜力的新型药物。
試驗對抗菌肽Dermadistinctin-Q1(DDQ1)進行原覈錶達,併對其生物活性進行檢測。採用重疊區擴增基因拼接法,多步PCR穫得抗菌肽DDQ1全基因,併使用原覈錶達載體pET-32a(+)構建工程菌。通過優化誘導錶達時間和異丙基硫代半乳糖苷(IPTG)終濃度,實現融閤蛋白錶達條件最佳化。利用腸激酶從融閤蛋白中分離抗菌肽DDQ1後,使用Ni離子親和層析法進行純化。進一步的生物活性檢測錶明,抗菌肽DDQ1具有廣譜抗菌活性和一定的熱、痠耐受性,是一類極具開髮潛力的新型藥物。
시험대항균태Dermadistinctin-Q1(DDQ1)진행원핵표체,병대기생물활성진행검측。채용중첩구확증기인병접법,다보PCR획득항균태DDQ1전기인,병사용원핵표체재체pET-32a(+)구건공정균。통과우화유도표체시간화이병기류대반유당감(IPTG)종농도,실현융합단백표체조건최가화。이용장격매종융합단백중분리항균태DDQ1후,사용Ni리자친화층석법진행순화。진일보적생물활성검측표명,항균태DDQ1구유엄보항균활성화일정적열、산내수성,시일류겁구개발잠력적신형약물。
This experiment expressed antibacterial peptide Dermadistinctin-Q1 (DDQ1)in prokaryotic system and de-tected its biological activity in vitro,the splice of genes by overlap extension was applied,and then the complete gene se-quence of DDQ1 was obtained by multistep PCR.The engineering bacteria was constructed by prokaryotic expression vector pET-32a (+).The optimization of fusion protein expression conditions came ture via optimizing the inducible expression time and the final concentration of IPTG.Antibacterial peptide DDQ1 was separated from the fusion protein by enterokinase and was purified by Ni ion affinity chromatography.The further detection of antibacterial peptide DDQ1 biological activitiy was performanced.The results showed that antibacterial peptide DDQ1 had broad-spectrum antibacterial activity,as well as a certain thermal-tolerance and acid-tolerance.Therefore,it was a kind of extremely potential new drug.