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A Journal on Biotechnology and Molecular Biology
Indexed/Abstracted in: EMBASE, Science Citation Index Expanded (SciSearch), Scopus
Impact Factor 0,246
Minerva Biotecnologica 2014 September;26(3):149-57
Using CM11 peptide as a cell permeable agent for the improvement of conventional plasmid transformation methods in Escherichia coli and Bacillus subtilis
Heiat M. 1, Aghamollaei H. 1, Moosazadeh Moghaddam M. 1, Kooshki H. 2
1 Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran;
2 Nanobiotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
AIM: Cold calcium chloride is the most practical and cheapest method for transformation of DNA into bacteria such as Escherichia coli, while some bacteria such as Bacillus subtilis uptake DNA in natural way. This study reports the increase of transformation efficiency using cationic peptide as a novel and efficient agent to transport DNA plasmids into Gram-negative and positive bacteria.
METHODS: pET-28a(+), pGEX4T-1 and pUC19 plasmids were separately transferred into E.coli, also B.subtilis was transformed by pWB980 plasmid. CaCl2 treated E.coli competent cells were prepared in the presence of different concentration of CM11 peptide in different time and mix with adequate amount of plasmids. In the case of B. subtilis, natural spontaneous DNA transformation accompanied with presence of different concentration of peptide.
RESULTS: In the presence of peptide, the transformation yield of pET-28a(+), pGEX4T-1 and pUC19 plasmids was 4.7, 4.4 and 4 fold higher than control, respectively. Results were also statistically significant p-value was less than 0.001 for all plasmids. Also the entry of pWB980 plasmid into B. subtilis with P<0.001 improved efficiently which was 6.4 fold higher than basal conditions.
CONCLUSION: These studies showed that CM11 peptide as a cell permeable peptide can increase the plasmid transformation efficiency in bacterial cells.