| dc.description.abstract | Transformation is a process used widely in molecular biology research and
biotechnology applications for gene cloning, protein expression, and genetic
engineering. However, transformation efficiency is generally low and influenced by
many factors, which must be improved to enhance cloning technology and reduce the
associated time, effort, and expense. Recent studies have shown that many factors can
affect transformation efficiency including ionic species, plasmid concentration, ice
bath time, and the recovery treatment time. Recently, cationic polymer has been
recognized as an effective transformation enhancer in increasing transformation
efficiency. In this study, we assessed the potential of two cationic poly(β-amino ester)
polymers, which were synthesized through an aza-Michael addition reaction between
piperazine and isosorbide diacrylate, as well as poly(ethylene glycol) diacrylate. These
polymers were investigated for their ability to enhance DNA transformation. Preparing
competent cells from E.coli strain DH5α was optimized by using the Ca2+ and adding
Mg2+
ions. To do it, different concentrations of the polymers were applied in pRSET EmGFP before transforming into competent cells through the heat shock method. The
colonies on the Ampicillin-added LB (Luria Bertani) agar were analyzed and
performed PCR reaction then the transformation efficiency was calculated. Based on
our findings, we observed a successful transformation of the emGFP gene, and the two
cationic poly(β-amino ester) polymers at high concentrations (20 - 100 ng/ul)
increased transformation efficiency more than twice as compared to normal
transformation. Notably, the polymer derived from isosorbide had the highest
transformation efficiency at a concentration of 20 ng/μl, while the poly(ethylene
glycol)-based polymer had lower transformation efficiency at low concentrations than the former. | en_US |
