星孢菌素产生菌Streptomyces sp.FIM18-327基因组的测序与遗传操作体系的建立

Genome sequencing of staurosporine-producing strain Streptomyces sp.FIM18-327 and establishment of its genetic manipulation system

目的通过基因组测序明确星孢菌素高产菌株Streptomyces sp.FIM18-327遗传背景,并建立该菌的遗传操作体系。方法通过第二代和第三代DNA测序技术相结合的策略对Streptomyces sp.FIM18-327进行全基因组测序并组装,利用antiSMASH软件预测该菌的次级代谢产物生物合成基因簇分布。通过优化接合转移载体、培养基种类及镁离子浓度,建立了Streptomyces sp.FIM18-327的遗传操作体系。结果Streptomyces sp.FIM18-327基因组大小为9.73 Mb,由一条染色体、两个线性质粒及一个环状质粒组成。其中染色体大小为9.48 Mb,GC含量70.12%,包含了8431个蛋白编码基因与34个次级代谢产物生物合成基因簇;以pFIM4123为载体,MS为接合转移培养基,在MS中添加MgCl2至终浓度为20 mmol/L时,接合转移的效率达3.2×10^(-6)。结论Streptomycessp.FIM18-327基因组信息的解析及遗传操作体系的建立,为该菌的进一步开发与应用奠定了基础。

Objective To elucidate the genetic background of the high-producing staurosporine strain Streptomyces sp.FIM18-327 and establish the genetic manipulation system for this strain.Methods The whole genome of Streptomyces sp.FIM18-327 was sequenced and assembled using both PacBio Sequel II and Illumina NovaSeq platforms,and the secondary metabolite biosynthesis gene clusters in the strain were predicted by antiSMASH.The genetic manipulation system of Streptomyces sp.FIM18-327 was established by investigating the conjugation transfer carrier,the type of culture medium and magnesium ion concentration.Results The genome size of this strain was 9.73 Mb,which consists of one chromosome,two linear plasmids and one circular plasmid.The chromosome size of Streptomyces sp.FIM18-327 was 9.48 Mb,with a GC content of 70.12%,containing 8,431 protein-coding genes and 34 secondary metabolite biosynthetic gene clusters.With pFIM4123 as the carrier,the efficiency of conjugation transfer reached 3.2×10^(-6),when MgCl_(2) was added to MS medium at the final concentration of 20 mmol/L.Conclusion The analysis of genomic information for Streptomyces sp.FIM18-327 and the establishment of a genetic manipulation system have laid the foundation for the further development and application of this strain.