摘要
为了更好地研究ε-聚赖氨酸(ε-PL)生物合成的分子机制以及构建ε-PL高产基因工程菌株,拟建立一株ε-PL产生菌株Streptomyces albulus PD-1的遗传转化体系。通过对培养基类型、孢子预萌发处理、培养基中Mg2+浓度等条件进行考察,以Escherichia coli ET12567(p UZ8002)为供体菌,成功地将p IB139质粒导入S.albulus PD-1中。结果表明:质粒转化效率达到(3±0.4)×10-6个接合转化子/受体。接合子传代实验和PCR结果发现,p IB139质粒能够稳定整合在S.albulus PD-1染色体的att B位点上。本研究建立了一株ε-PL生产菌株的遗传转化体系,为从分子水平上研究ε-PL的合成及ε-PL高产菌株的构建奠定了基础。
To study the biosynthetic mechanism of ε?Poly?l?lysine (ε?PL ) in the producing strain, we developed a genetic transformation system for Streptomyces albulus PD?1,a well?known ε?PL?producing strain. Plasmid pIB139 was introduced into S. albulus PD?1 when Escherichia coli ET12567( pUZ8002) was used as a donor. After optimizing for conjugal transfer conditions,a maximal conjugation frequency of (3?0±0?4) × 10?6 per recipient was obtained. Further test indicated that plasmid pIB139 was integrated into the attB site of S. albulus PD?1 chromosome,and the integration of plasmid pIB139 was stable after passing several generations. This inter?generic conjugal transformation system for S. albulus PD?1 will provide powerful tools to study ε?PL biosynthesis mechanism and to construct high ε?PL over?producers.
出处
《生物加工过程》
CAS
2016年第2期27-32,共6页
Chinese Journal of Bioprocess Engineering
基金
国家自然科学基金(21476112)
江苏省自然科学基金(BK20140933)
材料化学工程国家重点实验室自主课题(ZK201403)
