Preproabrin: genomic cloning, characterisation and the expression of the A-chain in Escherichia coli
3
Institute of Cancer Research, Royal Cancer Hospital, The Haddow Laboratories, Sutton, England
|
4
Genelabs Incorporated, Redwood City CA, USA
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Publication type: Journal Article
Publication date: 1991-06-01
scimago Q1
wos Q2
SJR: 2.212
CiteScore: 13.1
Impact factor: 4.2
ISSN: 1742464X, 00142956, 14321033, 17424658
PubMed ID:
2050149
Biochemistry
Abstract
Synthetic oligonucleotides representing all possible sequences of an N-terminal and an internal region of the A-chain of abrin C were used to generate a probe specific for abrin-related sequences using the polymerase chain reaction on Abrus precatorius genomic DNA. A lambda phage library constructed from genomic DNA isolated from leaf tissue of A. precatorius was screened and positive hybridising clones were characterised by restriction enzyme analysis. The coding regions of unique clones were characterised by DNA sequencing. One clone encodes a preproprotein closely related to abrin C with 83% similarity between the A-chain sequences. Based on similarity with the ricin toxins and Ricinus communis agglutinin, the preproabrin consists of an A-chain of 251 amino acids preceded by 34 amino acids containing an N-terminal signal peptide, followed by a 14-amino-acid linker and a B-chain of 263 amino acids. The mature A-chain of the preproabrin has been expressed cytoplasmically in Escherichia coli and the soluble recombinant protein was produced at levels exceeding 6% of total cell protein. The recombinant A-chain has been purified to homogeneity and its ability to depurinate 28S rRNA in rat liver ribosomes has been demonstrated in vitro.
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Citations from 2024:
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GOST
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Wood K. A. et al. Preproabrin: genomic cloning, characterisation and the expression of the A-chain in Escherichia coli // FEBS Journal. 1991. Vol. 198. No. 3. pp. 723-732.
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Wood K. A., LORD J. M., WAWRZYNCZAK E. J., Piatak M. Preproabrin: genomic cloning, characterisation and the expression of the A-chain in Escherichia coli // FEBS Journal. 1991. Vol. 198. No. 3. pp. 723-732.
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RIS
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TY - JOUR
DO - 10.1111/j.1432-1033.1991.tb16072.x
UR - https://doi.org/10.1111/j.1432-1033.1991.tb16072.x
TI - Preproabrin: genomic cloning, characterisation and the expression of the A-chain in Escherichia coli
T2 - FEBS Journal
AU - Wood, Katherine A.
AU - LORD, J. MICHAEL
AU - WAWRZYNCZAK, EDWARD J.
AU - Piatak, Michael
PY - 1991
DA - 1991/06/01
PB - Wiley
SP - 723-732
IS - 3
VL - 198
PMID - 2050149
SN - 1742-464X
SN - 0014-2956
SN - 1432-1033
SN - 1742-4658
ER -
Cite this
BibTex (up to 50 authors)
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@article{1991_Wood,
author = {Katherine A. Wood and J. MICHAEL LORD and EDWARD J. WAWRZYNCZAK and Michael Piatak},
title = {Preproabrin: genomic cloning, characterisation and the expression of the A-chain in Escherichia coli},
journal = {FEBS Journal},
year = {1991},
volume = {198},
publisher = {Wiley},
month = {jun},
url = {https://doi.org/10.1111/j.1432-1033.1991.tb16072.x},
number = {3},
pages = {723--732},
doi = {10.1111/j.1432-1033.1991.tb16072.x}
}
Cite this
MLA
Copy
Wood, Katherine A., et al. “Preproabrin: genomic cloning, characterisation and the expression of the A-chain in Escherichia coli.” FEBS Journal, vol. 198, no. 3, Jun. 1991, pp. 723-732. https://doi.org/10.1111/j.1432-1033.1991.tb16072.x.