Genetic recombination of Brevibacterium lactofermentum by protoplast fusion
1990
Lee, H.K. | Choi, S.Y. | Yoon, Y.K. | Min, K.H. (Sookmyung Women's Univ., Seoul (Korea Republic). Coll. of Sciences) | Rhee, Y.H. (Chungnam Univ., Taejon (Korea Republic). Dept. of Microbiology)
Brevibacterium lactofermentum SWA (arg trp) and B. lactofermentum SWB (met ser) were obtained from UV and NTG treatment. The rates of protoplast formation by B. lactofermentum SWA and SWB were 99.93 % and 99.98 %, respectively when each strain was treated with penicillin G in mid exponential growth phase, followed by incubation with 400 micro g/ml of lysozyme in lysis fluid supplemented with 0.4M sucrose. Frequencies of protoplast regeneration in B. lactofermentum SWA and B. lactofermentum SWB were 9.27 % and 10.32 % respectively, on regeneration medium containing 0.5M sodium succinate, 50 mM Mg** (2+), and 3 % PVP. In intraspecific protoplast fusion between B. lactofermentum SWA and B. lactofermentum SWB, fusion frequency of 2.30 x 10** (-5) was observed by using the 100 mM CaCl2 and 30 % PEG 6,000 in fusion fluid. Relative recombinant frequencies in each marker by means of selective media could be used for genetic analysis
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