한국미생물생명공학회:학술대회논문집 (Proceedings of the Korean Society for Applied Microbiology Conference)
- 한국미생물생명공학회 1986년도 추계학술대회
- /
- Pages.529.2-529
- /
- 1986
Expression of Tunicamycin Resistance in Bacillus subtilsls by Several Transfroming Plasmids
- Kong, In-Soo (Department of Food Engineering, Yonsei University) ;
- Makari-Yamasaki (Department of Agricultural Chemistry, The University of Tokyo)
- 발행 : 1986.12.01
초록
pSp-Si (1.6kbp) was originally found in pediococcus halophilus to be a cryptic multicopy-plasmid. Hoping that the plasmid can also replicate in Bacillus subtilis, protoplast transformation of strain 207-25 (recE) was performed using pSP-Sl onto which was added the marker of tmrB8 (on 4.9 kbp EcoRI fragment ) or tmrB+ (on 0.9 kbp xbaI fragment) gene. Though the tmrB8 gene can expres tunicamycin-resistance at the single copy state, and the tmrB+ gene exerts the resistance only at the multicopy state, we could not confirm the replication of pSP-Sl (tmrB8) or pSP-Sl(tmrB+) in B. subtilis. During the experiment, however, we unexpectedly found that the circularized 0.9 kbp xgaI fragment (tmrB+) itself, which had no replication origin, could transform strain 207-25 to tunicamycin-resistant by protoplast transformation. Southern hybridization analyses with tmrB+ and other probes revealed the integration of the fragment at a single copy state into a position other than the homologous tmrB gene. This recE independent integration of another tmrB+ gene into the chromosome may contribute to the tunicamycinresistance in the transformants.
키워드