Cloning and expression of heterologous genes in Rhodothermus marinus

Snaedis H. Bjornsdottir; Olafur H. Fridjonsson; Jakob K. Kristjansson; Gudmundur Eggertsson

doi:10.1007/s00792-006-0037-y

Cloning and expression of heterologous genes in Rhodothermus marinus

Snaedis H. Bjornsdottir, Olafur H. Fridjonsson, Jakob K. Kristjansson, Gudmundur Eggertsson^*

^*Corresponding author for this work

Faculty of Life and Environmental Sciences

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

7 Citations (Scopus)

Abstract

The construction of a shuttle cloning system for Rhodothermus marinus and Escherichia coli is described. It is based on the shuttle vector pRM3000, which contains a multiple cloning site as well as the shuttle marker trpB and TrpB^- recipients of both species. The vector is stable and in 25 ± 2 and 91 ± 3 copies in R. marinus SB-1 and E. coli SDH-1, respectively. Three different R. marinus regulatory sequences of the dnaJ, dnaK and recA genes were integrated into pRM3000 to make the expression vectors pRM5100, pRM5200 and pRM5300, respectively. Genes encoding α- and β-galactosidase (agaT and bgaT) from Thermus brockianus were cloned in R. marinus. Expression of both recombinant genes in R. marinus was demonstrated. The agaT gene was used as a reporter to study transcription from the expression vectors. Induced expression by ten- and twentyfold was observed from the dnaK and dnaJ regulatory sequences, respectively, after a temperature shift from 63 to 77°C. This is the first report of cloning and expression of heterologous genes in R. marinus and the first study of promoter activity in the species.

Original language	English
Pages (from-to)	283-293
Number of pages	11
Journal	Extremophiles
Volume	11
Issue number	2
DOIs	https://doi.org/10.1007/s00792-006-0037-y
Publication status	Published - Mar 2007

Bibliographical note

Funding Information:
Acknowledgments This work was supported by the Icelandic Research Centre. S.H. Bjornsdottir is a recipient of a grant from the Icelandic Research Fund for Graduate Students. O.H. Fridjonsson and J.K. Kristjansson also thank the EU 6 FP project DA-TAGENOM for support.

Other keywords

Expression vector
Reporter gene
Rhodothermus marinus
Shuttle vector
Thermo-induced expression

Access to Document

10.1007/s00792-006-0037-y

Cite this

@article{f4bbaa3e3bf0410d8e65698ad3a4464b,

title = "Cloning and expression of heterologous genes in Rhodothermus marinus",

abstract = "The construction of a shuttle cloning system for Rhodothermus marinus and Escherichia coli is described. It is based on the shuttle vector pRM3000, which contains a multiple cloning site as well as the shuttle marker trpB and TrpB- recipients of both species. The vector is stable and in 25 ± 2 and 91 ± 3 copies in R. marinus SB-1 and E. coli SDH-1, respectively. Three different R. marinus regulatory sequences of the dnaJ, dnaK and recA genes were integrated into pRM3000 to make the expression vectors pRM5100, pRM5200 and pRM5300, respectively. Genes encoding α- and β-galactosidase (agaT and bgaT) from Thermus brockianus were cloned in R. marinus. Expression of both recombinant genes in R. marinus was demonstrated. The agaT gene was used as a reporter to study transcription from the expression vectors. Induced expression by ten- and twentyfold was observed from the dnaK and dnaJ regulatory sequences, respectively, after a temperature shift from 63 to 77°C. This is the first report of cloning and expression of heterologous genes in R. marinus and the first study of promoter activity in the species.",

keywords = "Expression vector, Reporter gene, Rhodothermus marinus, Shuttle vector, Thermo-induced expression",

author = "Bjornsdottir, {Snaedis H.} and Fridjonsson, {Olafur H.} and Kristjansson, {Jakob K.} and Gudmundur Eggertsson",

note = "Funding Information: Acknowledgments This work was supported by the Icelandic Research Centre. S.H. Bjornsdottir is a recipient of a grant from the Icelandic Research Fund for Graduate Students. O.H. Fridjonsson and J.K. Kristjansson also thank the EU 6 FP project DA-TAGENOM for support.",

year = "2007",

month = mar,

doi = "10.1007/s00792-006-0037-y",

language = "English",

volume = "11",

pages = "283--293",

journal = "Extremophiles",

issn = "1431-0651",

publisher = "Springer",

number = "2",

}

TY - JOUR

T1 - Cloning and expression of heterologous genes in Rhodothermus marinus

AU - Bjornsdottir, Snaedis H.

AU - Fridjonsson, Olafur H.

AU - Kristjansson, Jakob K.

AU - Eggertsson, Gudmundur

N1 - Funding Information: Acknowledgments This work was supported by the Icelandic Research Centre. S.H. Bjornsdottir is a recipient of a grant from the Icelandic Research Fund for Graduate Students. O.H. Fridjonsson and J.K. Kristjansson also thank the EU 6 FP project DA-TAGENOM for support.

PY - 2007/3

Y1 - 2007/3

N2 - The construction of a shuttle cloning system for Rhodothermus marinus and Escherichia coli is described. It is based on the shuttle vector pRM3000, which contains a multiple cloning site as well as the shuttle marker trpB and TrpB- recipients of both species. The vector is stable and in 25 ± 2 and 91 ± 3 copies in R. marinus SB-1 and E. coli SDH-1, respectively. Three different R. marinus regulatory sequences of the dnaJ, dnaK and recA genes were integrated into pRM3000 to make the expression vectors pRM5100, pRM5200 and pRM5300, respectively. Genes encoding α- and β-galactosidase (agaT and bgaT) from Thermus brockianus were cloned in R. marinus. Expression of both recombinant genes in R. marinus was demonstrated. The agaT gene was used as a reporter to study transcription from the expression vectors. Induced expression by ten- and twentyfold was observed from the dnaK and dnaJ regulatory sequences, respectively, after a temperature shift from 63 to 77°C. This is the first report of cloning and expression of heterologous genes in R. marinus and the first study of promoter activity in the species.

AB - The construction of a shuttle cloning system for Rhodothermus marinus and Escherichia coli is described. It is based on the shuttle vector pRM3000, which contains a multiple cloning site as well as the shuttle marker trpB and TrpB- recipients of both species. The vector is stable and in 25 ± 2 and 91 ± 3 copies in R. marinus SB-1 and E. coli SDH-1, respectively. Three different R. marinus regulatory sequences of the dnaJ, dnaK and recA genes were integrated into pRM3000 to make the expression vectors pRM5100, pRM5200 and pRM5300, respectively. Genes encoding α- and β-galactosidase (agaT and bgaT) from Thermus brockianus were cloned in R. marinus. Expression of both recombinant genes in R. marinus was demonstrated. The agaT gene was used as a reporter to study transcription from the expression vectors. Induced expression by ten- and twentyfold was observed from the dnaK and dnaJ regulatory sequences, respectively, after a temperature shift from 63 to 77°C. This is the first report of cloning and expression of heterologous genes in R. marinus and the first study of promoter activity in the species.

KW - Expression vector

KW - Reporter gene

KW - Rhodothermus marinus

KW - Shuttle vector

KW - Thermo-induced expression

UR - http://www.scopus.com/inward/record.url?scp=33847259244&partnerID=8YFLogxK

U2 - 10.1007/s00792-006-0037-y

DO - 10.1007/s00792-006-0037-y

M3 - Article

C2 - 17124556

AN - SCOPUS:33847259244

SN - 1431-0651

VL - 11

SP - 283

EP - 293

JO - Extremophiles

JF - Extremophiles

IS - 2

ER -

Cloning and expression of heterologous genes in Rhodothermus marinus

Abstract

Bibliographical note

Other keywords

Access to Document

Fingerprint

Cite this