Transformation of Methanococcus maripaludis and identification of a Pst I-like restriction system

Debra L. Tumbula; Ronald A. Makula; William B. Whitman

doi:10.1111/j.1574-6968.1994.tb07118.x

Transformation of Methanococcus maripaludis and identification of a Pst I-like restriction system

Debra L. Tumbula, Ronald A. Makula, William B. Whitman

Research output: Contribution to journal › Article › peer-review

111 Scopus citations

Abstract

An optimized polyethylene glycol (PEG) method of transformation was developed for Methanococcus maripaludis using the pKAS102 integration vector. The frequency of transformation with 0.8 μg of plasmid and 3×10⁹ cells was 4.8×10^-5 transformants cfu^-1, or 1.8×10⁵ transformants μg^-1, which was four orders of magnitude greater than with the natural transformation method. A PstI restriction activity in M. maripaludis was also identified. Methylation of the plasmid with PstI methylase increased the methanococcal transformation frequency at least four-fold. Also, chromosomal DNA from M. maripaludis was resistant to digestion by the PstI endonuclease.

Original language	English (US)
Pages (from-to)	309-314
Number of pages	6
Journal	FEMS Microbiology Letters
Volume	121
Issue number	3
DOIs	https://doi.org/10.1111/j.1574-6968.1994.tb07118.x
State	Published - Sep 1 1994
Externally published	Yes

Keywords

Methanococcus maripaludis
Mrr
Polyethylene glycol
PstI
Restriction endonuclease
Transformation

ASJC Scopus subject areas

General Medicine

Access to Document

10.1111/j.1574-6968.1994.tb07118.x

Cite this

@article{d4fb7028e97d4e53aac4119798032a89,

title = "Transformation of Methanococcus maripaludis and identification of a Pst I-like restriction system",

abstract = "An optimized polyethylene glycol (PEG) method of transformation was developed for Methanococcus maripaludis using the pKAS102 integration vector. The frequency of transformation with 0.8 μg of plasmid and 3×109 cells was 4.8×10-5 transformants cfu-1, or 1.8×105 transformants μg-1, which was four orders of magnitude greater than with the natural transformation method. A PstI restriction activity in M. maripaludis was also identified. Methylation of the plasmid with PstI methylase increased the methanococcal transformation frequency at least four-fold. Also, chromosomal DNA from M. maripaludis was resistant to digestion by the PstI endonuclease.",

keywords = "Methanococcus maripaludis, Mrr, Polyethylene glycol, PstI, Restriction endonuclease, Transformation",

author = "Tumbula, {Debra L.} and Makula, {Ronald A.} and Whitman, {William B.}",

note = "Funding Information: We thank Jyoti Keswani for providing purified methanococcal chromosomal DNA. This work was supported by Department of Energy grant DE-FG09-91ER20045.",

year = "1994",

month = sep,

day = "1",

doi = "10.1111/j.1574-6968.1994.tb07118.x",

language = "English (US)",

volume = "121",

pages = "309--314",

journal = "FEMS Microbiology Letters",

issn = "0378-1097",

publisher = "Wiley-Blackwell",

number = "3",

}

TY - JOUR

T1 - Transformation of Methanococcus maripaludis and identification of a Pst I-like restriction system

AU - Tumbula, Debra L.

AU - Makula, Ronald A.

AU - Whitman, William B.

N1 - Funding Information: We thank Jyoti Keswani for providing purified methanococcal chromosomal DNA. This work was supported by Department of Energy grant DE-FG09-91ER20045.

PY - 1994/9/1

Y1 - 1994/9/1

N2 - An optimized polyethylene glycol (PEG) method of transformation was developed for Methanococcus maripaludis using the pKAS102 integration vector. The frequency of transformation with 0.8 μg of plasmid and 3×109 cells was 4.8×10-5 transformants cfu-1, or 1.8×105 transformants μg-1, which was four orders of magnitude greater than with the natural transformation method. A PstI restriction activity in M. maripaludis was also identified. Methylation of the plasmid with PstI methylase increased the methanococcal transformation frequency at least four-fold. Also, chromosomal DNA from M. maripaludis was resistant to digestion by the PstI endonuclease.

AB - An optimized polyethylene glycol (PEG) method of transformation was developed for Methanococcus maripaludis using the pKAS102 integration vector. The frequency of transformation with 0.8 μg of plasmid and 3×109 cells was 4.8×10-5 transformants cfu-1, or 1.8×105 transformants μg-1, which was four orders of magnitude greater than with the natural transformation method. A PstI restriction activity in M. maripaludis was also identified. Methylation of the plasmid with PstI methylase increased the methanococcal transformation frequency at least four-fold. Also, chromosomal DNA from M. maripaludis was resistant to digestion by the PstI endonuclease.

KW - Methanococcus maripaludis

KW - Mrr

KW - Polyethylene glycol

KW - PstI

KW - Restriction endonuclease

KW - Transformation

UR - http://www.scopus.com/inward/record.url?scp=0028168940&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028168940&partnerID=8YFLogxK

U2 - 10.1111/j.1574-6968.1994.tb07118.x

DO - 10.1111/j.1574-6968.1994.tb07118.x

M3 - Article

AN - SCOPUS:0028168940

SN - 0378-1097

VL - 121

SP - 309

EP - 314

JO - FEMS Microbiology Letters

JF - FEMS Microbiology Letters

IS - 3

ER -

Transformation of Methanococcus maripaludis and identification of a Pst I-like restriction system

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this