Kudriavtseva E A, Emel'ianova L K, Chinenova T A, Zvenigorodskiĭ V I, Voeĭkova T A
Genetika. 1996 Apr;32(4):510-6.
The level and the character of the Streptomyces virginiae virginiamycin-producing strain's resistance to self-produced antibiotic and a number of antibiotics from different groups were determined. S. virginiae was shown to display constitutive and inducible resistance to self-produced antibiotic. The phenomenon of cross-inducible resistance of the strain to virginiamycin and the antibiotics erythromycin, oleandomycin, and thiostrepton was demonstrated. The pTO1 and pVGTB24 plasmids were introduced into the strain by the method of intergeneric conjugation with Escherichia coli. Site-specific integration of the pTO1 vector into the S. virginiae chromosomal attB site without disturbance of growth, differentiation, and productivity of the strain was shown. The multicopy autonomously replicating plasmids pIJ699, pIJ702, pWOR109, and the integrative pZAT22 plasmid were introduced into the strain via electrotransformation of germinating spores. The efficiency of transformation was 1-5 x 10(3) transformants per 1 microgram DNA. The stable inheritance of the plasmids in S. virginiae without structural rearrangements was shown. These results allow the use of these plasmids to clone genes into S. virginiae.
测定了弗吉尼亚链霉菌(Streptomyces virginiae)产维吉尼亚霉素菌株对自身产生的抗生素以及多种不同类型抗生素的抗性水平和特性。结果表明,弗吉尼亚链霉菌对自身产生的抗生素表现出组成型和诱导型抗性。该菌株对维吉尼亚霉素以及红霉素、竹桃霉素和硫链丝菌肽等抗生素存在交叉诱导抗性现象。通过与大肠杆菌进行属间接合的方法,将pTO1和pVGTB24质粒导入该菌株。结果显示,pTO1载体可位点特异性整合到弗吉尼亚链霉菌染色体的attB位点,且不影响该菌株的生长、分化和生产力。通过对萌发孢子进行电转化,将多拷贝自主复制质粒pIJ699、pIJ702、pWOR109以及整合型pZAT22质粒导入该菌株。转化效率为每微克DNA产生1 - 5×10³个转化子。结果表明这些质粒在弗吉尼亚链霉菌中能够稳定遗传,且无结构重排。这些结果使得可以利用这些质粒将基因克隆到弗吉尼亚链霉菌中。