| 混合补料诱导高拷贝重组毕赤酵母高效表达X-HBsAg |
| |
| 引用本文： | 王龙刚,王宇凯,何成,楼觉人,杨忠东.混合补料诱导高拷贝重组毕赤酵母高效表达X-HBsAg[J].粉末涂料与涂装,2010,23(5). |
| |
| 作者姓名： | 王龙刚 王宇凯 何成 楼觉人 杨忠东 |
| |
| 作者单位： | 上海生物制品研究所,上海,200052 |
| |
| 摘 要： | 目的对高拷贝重组毕赤酵母表达X-HBsAg的诱导策略进行优化。方法在摇瓶中分别诱导1、4、8拷贝数的重组毕赤酵母,ELISA检测X-HBsAg的表达量;在15L发酵罐中对8拷贝的重组毕赤酵母进行诱导表达,分别考察甘油浓度对菌体比生长速率的影响、过渡阶段混合补料对菌体生长及细胞相对活性的影响、诱导阶段比生长速率对目的蛋白表达的影响以及诱导阶段混合补料对目的蛋白表达的影响。结果 8拷贝的重组毕赤酵母在摇瓶中诱导表达的X-HBsAg量明显高于1和4拷贝重组子。优化的发酵诱导策略为:初始补加甘油阶段控制甘油浓度在15g/L;过渡阶段采用甲醇与甘油混合补料,控制甘油浓度在限制性浓度以下;诱导阶段采用甲醇与甘油混合诱导X-HBsAg表达,诱导初始阶段控制比生长速率在0.015/h以上,中后期在0.002/h左右;发酵92h,X-HBsAg的表达量比单纯甲醇诱导提高约67%。结论混合补料诱导较单纯甲醇诱导策略更有利于高拷贝重组毕赤酵母表达X-HBsAg。
|
| 关 键 词： | 混合补料 X-HBsAg 高拷贝基因 毕赤酵母 发酵 |
Induction of High Expression of Multi-copy Recombinant X-HBsAg Gene in Pichia pastoris by Mixed Feeding |
| |
| WANG Long-gang,WANG Yu-kai,HE Cheng,et al.Induction of High Expression of Multi-copy Recombinant X-HBsAg Gene in Pichia pastoris by Mixed Feeding[J].Chinese Journal of Biologicals,2010,23(5). |
| |
| Authors: | WANG Long-gang WANG Yu-kai HE Cheng |
| |
| Affiliation: | WANG Long-gang,WANG Yu-kai,HE Cheng,et al(Shanghai Institute of Biological Products,Shanghai 200052,China) |
| |
| Abstract: | Objective To optimize the strategy for induction of high expression of multi-copy recombinant X-HBsAg gene in Pichia pastoris.Methods The recombinant P.pastoris with one-,four-and eight-copy X-HBsAg gene in shake-flask culture were induced,and the expression levels of X-HBsAg were determined by ELISA.The recombinant P.pastoris with eight-copy X-HBsAg gene was induced in 15 L fermentor,and the effects of glycerol concentration on specific growth rate of yeast,mixed feeding at transition phase on growth and r... |
| |
| Keywords: | X-HBsAg |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
