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FASCICOLI E ARTICOLI   I PIÙ LETTI   eTOC

ULTIMO FASCICOLOMINERVA BIOTECNOLOGICA

Rivista di Biologia Molecolare e Biotecnologie

Indexed/Abstracted in: EMBASE, Science Citation Index Expanded (SciSearch), Scopus
Impact Factor 0,246

Periodicità: Trimestrale

ISSN 1120-4826

Online ISSN 1827-160X

 

Minerva Biotecnologica 2014 Settembre;26(3):149-57

 ORIGINAL ARTICLES

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.

lingua: Inglese


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