摘要
旨在研究用乳糖替代IPTG作为诱导剂进行重组多价人精子抗原表位肽的表达及诱导表达的优化条件。在摇瓶发酵条件下,通过改变培养基组成、诱导时机、诱导温度、诱导剂浓度、诱导时间和诱导方式等条件,利用SDS-PAGE电泳和AlphaEase凝胶电泳图像分析系统,研究以上条件改变对GST-重组多价人精子抗原表位肽融合蛋白表达量的影响,并与IPTG诱导结果相比较。结果显示,在摇瓶试验中,最优表达条件为选用TB培养基,在菌体对数生长的中期进行诱导,诱导温度37℃、乳糖诱导终浓度3 mmol/L、诱导时间6 h。GST-重组多价人精子抗原表位肽融合蛋白的表达量占菌体总蛋白的30.5%,且主要以可溶形式表达,与IPTG的诱导结果相同。分批流加乳糖和一次性加入乳糖诱导,效果一样。乳糖可以替代IPTG作为诱导剂诱导GST-重组多价人精子抗原表位肽融合蛋白的表达,优化条件下可获得与IPTG相同的诱导表达效果。
It was to study lactose instead of IPTG as the inducer for expression of recombinant multi-epitopes peptide of human sperm antigens and optimal conditions of the expression.In the shake flask fermentation conditions,the influence of culture medium,induction period,induction temperature,inducer concentration,induction time and induction method on the expression level of GST-AG10-IR9-YLP12 fusion protein was analyzed with SDS-PAGE electrophoresis and AlphaEase gel electrophoresis image analysis system,and results were compared with that induced by IPTG.Results showed that obtained optimal conditions for the expression of GST-AG10-IR9-YLP12 fusion protein were as follows：TB as the medium,putting up induction in the middle logarithmic growth of bacteria,37℃ as induction temperature,3 mmol/L as the final induction concentration of lactose,extending induction for 6 h.The expressed fusion protein GST-AG10-IR9-YLP12 accounted for 30.5 percent of the bacterial total protein,mainly in a soluble form,and the expression level was the same as that induced by IPTG.The result of induced expression by adding lactose three times exhibited no different to that by adding lactose once.Therefore,it proved that lactose can substitute for IPTG as the inducer for expression of GST-AG10-IR9-YLP12 fusion protein,and the same induced expression level as that of IPTG can be obtained when use lactose to induce the fusion protein expression in optimal conditions.
出处
《生物技术通报》
CAS
CSCD
北大核心
2011年第7期171-175,共5页
Biotechnology Bulletin
基金
广西区科技厅科学研究与技术开发项目(桂科攻0632007-1I)
关键词
重组多价人精子抗原表位肽
乳糖
IPTG
表达
诱导
Recombinant multi-epitopes peptide of human sperm antigens Lactose IPTG Expression Induction
