English
 
Help Privacy Policy Disclaimer
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Cloning vectors for the production of proteins in Dictyostelium discoideum

Manstein, D. J., Schuster, H.-P., Morandini, P., & Hunt, D. M. (1995). Cloning vectors for the production of proteins in Dictyostelium discoideum. Gene, 162(1), 129-134. doi:10.1016/0378-1119(95)00351-6.

Item is

Basic

show hide
Genre: Journal Article
Alternative Title : Cloning vectors for the production of proteins in Dictyostelium discoideum

Files

show Files
hide Files
:
Gene_162_1995_129.pdf (Any fulltext), 526KB
 
File Permalink:
-
Name:
Gene_162_1995_129.pdf
Description:
-
Visibility:
Restricted (Max Planck Institute for Medical Research, MHMF; )
MIME-Type / Checksum:
application/pdf
Technical Metadata:
Copyright Date:
-
Copyright Info:
-
License:
-

Creators

show
hide
 Creators:
Manstein, Dietmar J.1, 2, Author              
Schuster, Hans-Peter, Author
Morandini, Piero, Author
Hunt, Deborah M., Author
Affiliations:
1Dietmar Manstein Group, Max Planck Institute for Medical Research, Max Planck Society, ou_1497708              
2Emeritus Group Biophysics, Max Planck Institute for Medical Research, Max Planck Society, ou_1497712              

Content

show
hide
Free keywords: Recombinant DNA; myosin; cytoskeleton; expression vector; cellular slime mold; transformation
 Abstract: We constructed and tested a series of cloning vectors designed to facilitate protein production and purification in Dictyostelium discoideum (Dd). These vectors carry the origin of replication of the Dd high-copy-number plasmid Ddp2, expression cassettes consisting of the strong, constitutive actin (act15) or the inducible discoidin (disI gamma) promoters, a translational start codon upstream from a multiple cloning site and sequences for the addition of epitope or affinity tags at the N- or C-termini of any protein. The affinity tag used corresponds to 7 (N-terminal fusion) or 8 (C-terminal fusion) His residues. The epitope tags correspond to an 11-amino-acid sequence from human c-myc, recognised by monoclonal antibody (mAb) 9E10, and the Glu-Glu-Phe sequence recognised by mAb YL1/2 to alpha-tubulin. Both these mAb are commercially available. The YL1/2 epitope offers a second affinity tag for the purification of proteins under native conditions. The functional competence of the vectors was tested by determining their ability to promote the expression of various Dd myosin constructs. High synthesis levels were obtained for each vector; up to 1 mg of homogenous, functional protein per g of cells was obtained after purification of the recombinant products.

Details

show
hide
Language(s): eng - English
 Dates: 1995-02-011995-03-151995-08-30
 Publication Status: Published in print
 Pages: 6
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Gene
  Other : Gene
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: Amsterdam : Elsevier
Pages: - Volume / Issue: 162 (1) Sequence Number: - Start / End Page: 129 - 134 Identifier: ISSN: 0378-1119
CoNE: https://pure.mpg.de/cone/journals/resource/954925526821