Predictive mutagenesis of ligation-independent cloning (LIC) vectors for protein expression and site-specific chemical conjugation

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Standard

Predictive mutagenesis of ligation-independent cloning (LIC) vectors for protein expression and site-specific chemical conjugation. / Vernet, Erik; Sauer, Jørgen; Andersen, Peter Andreas; Jensen, Knud Jørgen; Voldborg, Bjørn Gunnar Rude.

I: Analytical Biochemistry, Bind 414, Nr. 2, 2011, s. 312-314.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt

Harvard

Vernet, E, Sauer, J, Andersen, PA, Jensen, KJ & Voldborg, BGR 2011, 'Predictive mutagenesis of ligation-independent cloning (LIC) vectors for protein expression and site-specific chemical conjugation', Analytical Biochemistry, bind 414, nr. 2, s. 312-314. https://doi.org/10.1016/j.ab.2011.03.015

APA

Vernet, E., Sauer, J., Andersen, P. A., Jensen, K. J., & Voldborg, B. G. R. (2011). Predictive mutagenesis of ligation-independent cloning (LIC) vectors for protein expression and site-specific chemical conjugation. Analytical Biochemistry, 414(2), 312-314. https://doi.org/10.1016/j.ab.2011.03.015

Vancouver

Vernet E, Sauer J, Andersen PA, Jensen KJ, Voldborg BGR. Predictive mutagenesis of ligation-independent cloning (LIC) vectors for protein expression and site-specific chemical conjugation. Analytical Biochemistry. 2011;414(2):312-314. https://doi.org/10.1016/j.ab.2011.03.015

Author

Vernet, Erik ; Sauer, Jørgen ; Andersen, Peter Andreas ; Jensen, Knud Jørgen ; Voldborg, Bjørn Gunnar Rude. / Predictive mutagenesis of ligation-independent cloning (LIC) vectors for protein expression and site-specific chemical conjugation. I: Analytical Biochemistry. 2011 ; Bind 414, Nr. 2. s. 312-314.

Bibtex

@article{823629b2913949c394fff004b025aa1a,

title = "Predictive mutagenesis of ligation-independent cloning (LIC) vectors for protein expression and site-specific chemical conjugation",

abstract = "Ligation-independent cloning (LIC) allows for cloning of DNA constructs independent of insert restriction sites and ligases. However, any required mutations are typically introduced by additional, time-consuming steps. We present a rapid, inexpensive method for mutagenesis in the 5' LIC site of expression constructs and report on the construction of expression vectors with N-terminal serine, cysteine, threonine, or tyrosine residues after tobacco etch virus (TEV) protease cleavage. In a practical application, the N-terminal serine was oxidized to an aldehyde, subsequently reacted with an amino-oxy functionalized polyethylene glycol (PEG) ligand under aniline catalysis to provide a protein selectively modified at the N-terminus.",

keywords = "Amino Acid Sequence, Cloning, Molecular, Endopeptidases, Genetic Vectors, Molecular Sequence Data, Mutagenesis, Plasmids, Polyethylene Glycols, Recombinant Proteins, Serine",

author = "Erik Vernet and J{\o}rgen Sauer and Andersen, {Peter Andreas} and Jensen, {Knud J{\o}rgen} and Voldborg, {Bj{\o}rn Gunnar Rude}",

year = "2011",

doi = "10.1016/j.ab.2011.03.015",

language = "English",

volume = "414",

pages = "312--314",

journal = "Analytical Biochemistry",

issn = "0003-2697",

publisher = "Elsevier",

number = "2",

}

RIS

TY - JOUR

T1 - Predictive mutagenesis of ligation-independent cloning (LIC) vectors for protein expression and site-specific chemical conjugation

AU - Vernet, Erik

AU - Sauer, Jørgen

AU - Andersen, Peter Andreas

AU - Jensen, Knud Jørgen

AU - Voldborg, Bjørn Gunnar Rude

PY - 2011

Y1 - 2011

N2 - Ligation-independent cloning (LIC) allows for cloning of DNA constructs independent of insert restriction sites and ligases. However, any required mutations are typically introduced by additional, time-consuming steps. We present a rapid, inexpensive method for mutagenesis in the 5' LIC site of expression constructs and report on the construction of expression vectors with N-terminal serine, cysteine, threonine, or tyrosine residues after tobacco etch virus (TEV) protease cleavage. In a practical application, the N-terminal serine was oxidized to an aldehyde, subsequently reacted with an amino-oxy functionalized polyethylene glycol (PEG) ligand under aniline catalysis to provide a protein selectively modified at the N-terminus.

AB - Ligation-independent cloning (LIC) allows for cloning of DNA constructs independent of insert restriction sites and ligases. However, any required mutations are typically introduced by additional, time-consuming steps. We present a rapid, inexpensive method for mutagenesis in the 5' LIC site of expression constructs and report on the construction of expression vectors with N-terminal serine, cysteine, threonine, or tyrosine residues after tobacco etch virus (TEV) protease cleavage. In a practical application, the N-terminal serine was oxidized to an aldehyde, subsequently reacted with an amino-oxy functionalized polyethylene glycol (PEG) ligand under aniline catalysis to provide a protein selectively modified at the N-terminus.

KW - Amino Acid Sequence

KW - Cloning, Molecular

KW - Endopeptidases

KW - Genetic Vectors

KW - Molecular Sequence Data

KW - Mutagenesis

KW - Plasmids

KW - Polyethylene Glycols

KW - Recombinant Proteins

KW - Serine

U2 - 10.1016/j.ab.2011.03.015

DO - 10.1016/j.ab.2011.03.015

M3 - Journal article

C2 - 21414287

VL - 414

SP - 312

EP - 314

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 2

ER -

ID: 40289329

Kemisk Institut