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PMID: 3116931 Published · ppublish English Journal Article

A method for genetic transformation of nonprotoplasted Streptococcus lactis.

Applied and environmental microbiology ·Vol. 53 ·No. 8 ·1987-08-00 ·Pages 1730-6

Sanders ME, Nicholson MA

Abstract

Plasmid transformation of whole cells of Streptococcus lactis LM0230 was demonstrated. The procedure required polyethylene glycol and incubation in hypertonic media, but did not require enzymatic cell wall digestion. Conditions were optimized, yielding 5 X 10(5) transformants per micrograms of pSA3 DNA. Variables tested for effect on transformation efficiency included molecular weight, concentration, and pH of polyethylene glycol; cell density; plating media; DNA concentration; heat shock; and incubation of cells in hypertonic buffer. DNAs transformed included pSA3, pVA856, pTV1, and c2 phi. Transformation from DNA-DNA ligation mixes, with DNA not purified through density gradients, and with previously frozen cells was also achieved. The method described here for transformation of nonprotoplasted cells of LM0230 is unique, and to date has not been applied successfully to other lactic acid bacteria.

MeSH Terms
Cloning, Molecular Culture Media DNA, Bacterial/analysis,genetics Electrophoresis, Agar Gel Hydrogen-Ion Concentration Lactococcus lactis/genetics Molecular Weight Nucleic Acid Hybridization Plasmids Polyethylene Glycols Transformation, Bacterial
Chemicals
Culture Media DNA, Bacterial Polyethylene Glycols
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Sanders M E
Biotechnology Group, Marschall/Miles Laboratories, Elkhart, Indiana 46515.
Nicholson M A
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20 references, click to expand
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/aem.53.8.1730-1736.1987
Published
1987-08-00
Pages
1730-6
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC203982
Subset
IM
External Links
PubMed source DOI: 10.1128/aem.53.8.1730-1736.1987 PMC Full Text View journal details
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