Dam and Dcm methylations prevent gene transfer into Clostridium pasteurianum NRRL B-598: development of methods for electrotransformation, conjugation, and sonoporation

Table 4 Summary of pMTL83353 containing CFU yielded by conjugation, sonoporation, and combined sono/electroporation approaches

Method	CFU (average count)	Efficiency (CFU per μg DNA)
Conjugation (E. coli JM110 containing RP4 and pMTL83253 donor)
6 h of conjugation	12
24 h of conjugation	37
Sonoporation
10 % PEG 8000 buffer, 20 s pulse	225^a	112.5
30 % PEG 8000 buffer, 20 s pulse	321^a	160.5
Sono/electroporation
30 % PEG 8000 buffer, 20 s ultrasound pulse, 5 ms square-wave pulse 5 ms (1250 V)	265^b	530

ISSN: 2731-3654