Baltz R H
J Gen Microbiol. 1978 Jul;107(1):93-102. doi: 10.1099/00221287-107-1-93.
Conditions for highly efficient genetic recombination in Streptomyces by protoplast fusion are described. Protoplasts of S. fradiae and S. griseofuscus were formed by a modification of the glycine-lysozyme-lytic enzyme method (Okanishi, Suzuki & Umezawa, 1974). Regeneration of cells from protoplasts was monitored throughout the growth cycle and was most efficient when cells of either S. fradiae or S. griseofuscus were taken from the transition phase between the exponential and stationary growth phases. Fusion of protoplasts carrying different auxotrophic or chromosomal drug-resistance markers was achieved by treatment with polyethylene glycol, and high frequencies of stable genetic recombinants were obtained.
描述了通过原生质体融合在链霉菌中进行高效基因重组的条件。通过改良甘氨酸 - 溶菌酶 - 裂解酶法（冈西、铃木和梅泽，1974年）制备了弗氏链霉菌和灰褐链霉菌的原生质体。在整个生长周期中监测原生质体细胞的再生情况，当弗氏链霉菌或灰褐链霉菌的细胞取自指数生长期和稳定生长期之间的过渡阶段时，再生效率最高。通过聚乙二醇处理实现了携带不同营养缺陷型或染色体抗药标记的原生质体融合，并获得了高频率的稳定基因重组体。
