원문정보
초록
영어
Propionibacterium acnes has been known to be involved in the pathology of acnes. However, the definite mechanism in development of acnes and the inflammation are unknown. For P. acnes transformation method has not been established, although it is believed to be basic tool for gene manipulation. In this study we had attempted to develop P. acnes transformation method by using
electroporation. Various parameters were used to develop and optimize the transformation of P. acnes. Among them two factors were crucial in the transformation for P. acnes: one was the E. coli strain from which the plasmid DNA had been isolated and the other was the growth temperature of P. acnes competent cells. It was essential to prepare plasmid DNA from a dam- E. coli strain, ET12567. When plasmid DNAs isolated from the other E. coli strains
such as JM109 and HB101 were tested, transformation efficiency was extremely low. When P. acnes cells were cultivated at 24℃for competent cell preparation, transformation efficiency was increased considerably. When plasmid DNA isolated from a dam- mutant strain of E. coli was used for transformation of P. acnes which had been grown at 24℃ maximum transformation efficiency of 1.8×104 transformants per ㎍ of plasmid DNA was obtained at field strength of
6kV/cm with pulse time at 3.2 ms. This is believed to be the first report on the transformation of P. acnes which can be employed for gene manipulations including knock-out of specific genes.