摘要
对蜂毒素基因进行改造后,通过PCR方法获得新蜂毒素基因(MEA),将其克隆到表达载体pPICZa-A,而后将重组表达载体pPICZa-A-MEA转化GS115,筛选获得重组酵母菌.对GS115-ME经甲醇诱导表达并对培养条件进行优化探讨.对改造的蜂毒素进行了溶血活性、热稳定性及酸碱稳定性测定.结果表明,蜂毒素基因成功地在毕赤酵母中表达,经改造的蜂毒素在保留了抗菌活性的同时溶血活性降低20倍左右,同时还具有良好的热稳定性和酸碱稳定性.
In order to reduce hemolysis activity extremely and keep antibacter ia l activity of melittin,the amino acid sequence of melittin was altered by geneti c engineering.Then,the new melittin gene has been expressed in Pichia pastoris.a nd the positive clones were selected by antibiotics.After yeast cell culture con ditions had been optimized,the positive clones were induced with methanol to exp ress melittin.The results showed that the hemolysis activity of melittin was gre atly depressed and it still keeped substantid antibacterial activity.
出处
《生命科学研究》
CAS
CSCD
2005年第2期124-128,共5页
Life Science Research
基金
广东省自然科学基金资助项目(980174)
关键词
蜂毒素基因
毕赤酵母
分子改造
条件优化
溶血活性
melittin
Pichia pastoris
molecular designing
culture conditions
hem olysis activity
