抗生素Bagremycin生产菌Streptomyces sp. Tü4128接合转移系统的建立

Development of an Intergeneric Conjugal Transfer System for Streptomyces sp. Tü 4128,a Producer of Bagremycins

基于噬菌体φC31 att/int系统,采用属间接合将来自大肠杆菌的DNA有效地导入菌株Tü4128的孢子中。通过条件优化,当融合温度为37℃时,在含80mmol/L MgCl2的ISP4培养基上使每个受体细胞的接合频率显著提高到7.3×10-3。Southern blot分析显示在受体菌染色体上存在单一的染色体附着位点(attB位点)。整合位点的DNA序列测序显示为保守。采用优化的方法建立了大肠杆菌和菌株Tü4128间高效的属间接合系统。

DNA from Escherichia coli was successfully introduced into spores of strain Tü 4128 by intergeneric conjugation through bacteriophage φC31 att/int system.The transconjugation frequency on ISP4 medium with 80 mmol/L MgCl2 was significantly improved up to 7.3×10-3 per recipient cell under optimized mating temperature of 37 ℃.Southern blot analysis revealed a single chromosomal attachment site on the chromosome of the recipient strain.The DNA sequence of the integration site was determined and shown to be conserved.An efficient intergeneric conjugation between Escherichia coli and strain Tü 4128 was successfully achieved by condition optimization.