摘要
重组大肠杆菌BL21携带抗克伦特罗单链抗体(scFv)基因,研究了不同培养基、诱导时期、诱导时间、初始pH以及装液量对重组抗体表达量的影响,并对重组质粒稳定性进行研究。结果表明,在初始pH为7.2、装液量为25mL/250mL的LB培养基中,30℃培养菌体生长至A600nm为0.8时,42℃诱导表达6h,scFv表达量可达33.7%。工程菌在无选择压力条件下连续传代120代,保持稳定,100%的菌株携带重组质粒且经诱导后均有重组抗体的表达。
The recombinant E.coil BL21 carried anti-clenbuterol single chain fragment variable gene.Different factors influencing expression of recombinant scFv antibody such as medium, inducing period, inducing time, initial pH and medium volume were investigated.The results showed under optimal conditions, namely the bacteria density was A600nm 0.8 in LB medium,inducing time 6h, inducing temperature 42℃,initial pH ？.2, medium volume 25ml_/250mL flask,the expression level reaches up to 33.7% of the total bacteria protein.Also, the recombinant plasmid was stable after cultivation in LB medium at 30℃ for 120 generations without antibiotic selection.
出处
《食品工业科技》
CAS
CSCD
北大核心
2009年第7期175-177,共3页
Science and Technology of Food Industry
基金
"863"计划(2007AA10Z437)
国家自然科学基金(30871755
30400354)
高等学校博士点基金(20050564011)
广东省自然科学基金(06025825)
粤港关键领域重点突破招标项目(2006A25005002)
关键词
克伦特罗
单链抗体
工程菌
培养条件
clentuberol
single chain Fv
engineered strain
culture conditions
