Genetic Recombination of Brevibacterium lactofermentum by Protoplast Fusion

Brevibacterium lactofermentum의 원형질체 융합에 의한 유전자 재조합

  • 이혜경 (숙명여자대학교 이과대학 생물학과) ;
  • 최순영 (숙명여자대학교 이과대학 생물학과) ;
  • 윤윤경 (숙명여자대학교 이과대학 생물학과) ;
  • 이영하 (충남대학교 미생물학과) ;
  • 민경희 (숙명여자대학교 이과대학 생물학과)
  • Published : 1990.06.01

Abstract

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 $\mu\textrm{g}$/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 $2.30\times 10^{-5}$ was observed by using the 100mM $CaCl_{2}$ 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.

Keywords

Protoplast fusion;ggnetic anallysis of recombinasts Brevibacterium lactofermentum