摘要
对东方拟无枝酸菌HCCB10007原生质体制备和再生条件进行了探索,结果表明以含2%甘氨酸的TSB培养基培养菌丝体,28℃经1mg/ml溶菌酶处理30min,形成的原生质体约达107个/ml,原生质体再生率为2.5%。将来源于HCCB10007的gtfE基因通过安普霉素抗性基因插入失活,构建质粒pLY26并经PEG介导转化HCCB10007原生质体,PCR鉴定结果表明转化成功。
The conditions for protoplasts preparation and regeneration from Amycolatopsis orientalis HCCB10007 were optimized. The results showed that about 107/ml of protoplasts was formed, with regeneration frequency of 2.5%, when the mycelia were incubated in TSB medium containing 2% glycine and treated with 1mg/ml lysozyme for 30 min at 28℃. PCR Identification gave the conclusion that the plasmid pLY26, containing the disrupted gtfE from the chromosomal DNA of A. orientalis HCCB10007 by inserting apramycin resistance marker gene, was introduced into A. orientalis HCCB10007 by PEG-assisted transformation of protoplasts.
出处
《中国医药工业杂志》
CAS
CSCD
北大核心
2005年第6期332-335,共4页
Chinese Journal of Pharmaceuticals
