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为了提高GABAA受体a1蛋白片段在大肠杆菌中表达量,研究了重组菌的发酵条件,包括培养基、接种量、温度、摇床转速、pH、诱导培养时间和诱导剂IPTG使用浓度等对GABAA受体蛋白片段表达的影响。结果表明重组菌以LB培养基为发酵基质,按3%接种量,37℃培养细胞3.5h后IPTG 32℃诱导5h,菌体生物量为3.25g/L,目标蛋白表达量达95mg/L。用16L发酵罐进行放大培养,菌体生物量达4.95g/L,发酵周期5.5h,最高目标蛋白表达量达到136mg/L. 相似文献
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7号淀粉酶链霉菌M1033木糖异构酶基因的启动子活性检测、转录起始点的确定 总被引:1,自引:0,他引:1
分别从重组质粒pUB1及其M(13)亚克隆S1中将7号淀粉酶链霉菌(StreptomycesdiastaticusNo.7)M1033(以下简称S.di.M1033)木糖异构酶基因的-192-+581bp片段克隆入链霉菌启动子探测质粒pIJ4083中,转化变铅青链霉菌(S.lividans)TK24。通过对其邻苯二酚加双氧酶活性的检测表明,该片段具有启动子活性。应用M13亚克隆S1和合成引物P6延伸制备放射性标记的单链DNA探针;通过S.di.M1033的总RNA的S1核酸酶保护实验,确定了其转录的起始位点,并由此探讨了与木糖异构酶基因表达有关的一些因素。 相似文献
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影响白腐真菌AH28-2菌株漆酶合成的因子和发酵条件 总被引:13,自引:0,他引:13
White-rot fungus AH28-2, a newly isolated strain, produced effectively laccase by induction when grown on a synthetic medium. Aromatic compounds of low molecular weight had an inducing influence on laccase production and its isoenzyme compositions. The using of o-toluidine or syringic acid had the best inducing effect. Cu2+ concentration in medium had distinguished effect on laccase production. Enzyme activity was notably increased by Cu2+ and reached the maximum when Cu2+ final concentration was 5 mumol/L. Mn2+ inhibited the synthesis of laccase. Carbon and nitrogen limitation were not beneficial to laccase synthesis, while high nutrient organic medium was beneficial to the growth of cell and the synthesis of laccase. Using cellobiose as the sole carbon source, the highest level enzyme activity reached 82,923. 7 u/L under the condition of optimum fermentation with ABTS as substrate. This enzyme activity was 2.9-fold higher compared to the reported data on international references in recent years. 相似文献
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分别从重组质粒PUB1及其M13亚克隆S1中将7号淀粉酶链霉菌(Strcptomyces diastaticus No.7)M1033(以下简称S。di.M1033)木糖异构酶基因的-192-+581bp片段克隆入链霉菌启动子探测质粒PIJ4083中,转化变铅青链霉菌(S。lividans)TK24。通过对其邻苯二酚加双氧酶活性的检测表明,该片段具有启动子活性。应用M13亚克隆S1和合成引物P6延 相似文献
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