摘要
目的通过对阿维菌素生物合成基因C5-O-甲基转移酶基因(aveD)内部进行缺失,使aveD基因失活,从而获得只产生阿维菌素B组分的基因工程菌。方法构建大肠杆菌-链霉菌重组质粒pZHJ06,通过接合转移将缺失部分片段的aveD基因以双交换的方式整合到阿维链霉菌的染色体上。采用摇瓶进行发酵,采用高效液相色谱(HPLC)检测发酵液中阿维菌素的组分。结果重组菌株不再产生阿维菌素A组分,只产生阿维菌素B组分,并且B组分的总产量也有所提高。结论将阿维链霉菌的aveD基因失活,并不影响下游酮基还原酶基因(aveF)基因的表达,重组菌株只产生阿维菌素B组分。
Objective To construct a gene-engineered strain which producing avermectin B components only by inactivating aveD gene in the chromosome.Methods An E.coli-Streptomyces shuttle plasmid pZHJ06 containing inactive aveD gene was constructed and was introduced into Streptomyces avermitilis through conjugal transfer.Then aveD gene in the chromosome was replaced by the inactive aveD gene in the plasmid pZHJ06.Avermectin fermentation was conducted by shaking flask method and avermectin components was determined by HPLC method.Results The recombinant strain produced avermectin B components only,avermectin A components were disappeared.Meanwhile,the total yield of avermectin B was also increased.Conclusions Inactivating aveD did not affect the expression of aveF downstream of aveD gene.The recombinant strain produced avermectin B components only.
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2010年第11期919-923,共5页
Journal of Shenyang Pharmaceutical University
作者简介
周海娇（1985-），女（汉族），黑龙江五常人，硕士研究生，E-mail：zhouhaijiao85@163．com
何建勇（1952-），男（汉族），天津人，教授，主要从事微生物制药及链霉菌分子生物学研究，Tel．024-23986401，E-mail jianyonghe@sina．com
