正交试验法在优化重组乳酸乳球菌诱导表达中的应用研究
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摘要
为提高重组乳酸菌NICE诱导表达系统表达外源蛋白的能力,研究选取Nisin加入量、OD值、诱导温度和诱导时间这4个主要影响表达的因素,每个因素设置3个水平,设计4因素3水平(43)正交试验。通过优化Nisin加入量、OD值、诱导温度和诱导时间,从而确定重组乳酸菌表达外源蛋白最佳条件。结果显示:当Nisin的量为1 ng/mL、OD值为0.3、诱导温度为30℃、诱导时间为7 h时,其重组乳酸菌表达外源蛋白的量最高。研究结果为由Nisin控制表达基因系统的重组乳酸乳球菌疫苗开发奠定了基础。
In order to improve the ability of recombinant lactic acid bacteria NICE induced expression system to express foreign proteins, this study selected three levels from each factor based on the four main factors affecting the expression of Nisin supplementation, OD value, induction temperature and induction time. The orthogonal experimental table was designed for 4 factors and 3 levels. By optimizing the three levels of Nisin supplementation, OD value,induction temperature and induction time, the optimal conditions for the expression of exogenous proteins in recombinant lactic acid bacteria were determined. The results showed that when the amount of Nisin was 1 ng/mL, OD value was0.3, the induction temperature was 30 ℃ and the induction time was 7 h, the recombinant lactic acid bacteria expressed the highest amount of foreign protein. This study laid the foundation for the vaccine development of recombinant Lactococcus lactis with the gene expression system controled by Nisin.
In order to improve the ability of recombinant lactic acid bacteria NICE induced expression system to express foreign proteins, this study selected three levels from each factor based on the four main factors affecting the expression of Nisin supplementation, OD value, induction temperature and induction time. The orthogonal experimental table was designed for 4 factors and 3 levels. By optimizing the three levels of Nisin supplementation, OD value,induction temperature and induction time, the optimal conditions for the expression of exogenous proteins in recombinant lactic acid bacteria were determined. The results showed that when the amount of Nisin was 1 ng/mL, OD value was0.3, the induction temperature was 30 ℃ and the induction time was 7 h, the recombinant lactic acid bacteria expressed the highest amount of foreign protein. This study laid the foundation for the vaccine development of recombinant Lactococcus lactis with the gene expression system controled by Nisin.
引文
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[9]刘力章,黄锦勇,吴春芳.正交试验法在固定化微生物技术处理乳品废水工艺条件优化中的应用[J].江西科学,2010,28(6):772-774.
[10]楚遵锋,徐淑华,王敏,等.乳酸杆菌基因工程载体的应用[J].今日畜牧兽医,2017(1):49-51.
[2] LIU L, ZHANG W, SONG Y, et al. Recombinant Lactococcus lactis, co-expressing OmpH of an M cell-targeting ligand and IBDV-VP2 protein provide immunological protection in chickens[J]. Vaccine, 2017, 36(5):729-735.
[3]徐波.食品级高效诱导表达系统-NICE系统[J].生物技术通报,2005(2):14-17.
[4]张晶晶,李泰明,谭树华.乳酸菌食品级nisin控制的基因表达系统NICE[J].中国生物工程杂志,2006(3):68-72.
[5]王玄静.正交试验设计的应用及分析[J].兰州文理学院学报(自然科学版),2016,30(1):17-22.
[6] KANAYAMA M, KATO Y, TSUJI T, et al. Enhancement of immunomodulative effect of lactic acid bacteria on plasmacytoid dendritic cells with sucrose palmitate[J]. Scientific Reports, 2018, 8(1):3147.
[7] MAYS Z J, NAIR N U. Synthetic biology in probiotic lactic acid bacteria:At the frontier of living therapeutics[J].Current Opinion in Biotechnology, 2018(53):224-231.
[8] PERCZAK A, GOLINSKI P, BRY?A M, et al. The efficiency of lactic acid bacteria against pathogenic fungi and mycotoxins[J]. Arhiv Za Higijenu RadaⅠToksikologiju, 2018, 69(1):32.
[9]刘力章,黄锦勇,吴春芳.正交试验法在固定化微生物技术处理乳品废水工艺条件优化中的应用[J].江西科学,2010,28(6):772-774.
[10]楚遵锋,徐淑华,王敏,等.乳酸杆菌基因工程载体的应用[J].今日畜牧兽医,2017(1):49-51.