Linked Life Data

2045456

Source:http://linkedlifedata.com/resource/pubmed/id/2045456

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1991-7-18
pubmed:abstractText
The cloning of a cDNA coding for the skeletal human beta-tropomyosin in the bacterial expression vector pKK233-2 is reported. Deletion mutants were also constructed. pCF-T1088 was obtained by elimination of exon 9 and pCF-T1089 was built by deleting 2/3 of the first exon. The recombinant tropomyosins were synthesized in E. coli after induction by IPTG. The mutant proteins were characterized by western blot using antibodies raised against native tropomyosin. The amount of the human protein synthesized in E. coli varies with each mutant, suggesting the involvement of the structure of the protein or of the mRNA on the synthesis or the stability of the recombinant protein. After precipitation of most of the bacterial proteins at 100 degrees C, purification was achieved by high-performance liquid chromatography (HPLC) using TSK-DEAE, hydroxyapatite and reversed-phase columns. The chromatographic behaviour of the mutants were compared. Characterization of the mutated tropomyosins was achieved by tryptic digestion and analysis of the peptide composition by reversed-phase HPLC. A computer program for predicting the retention times of the peptides generated was written. It is shown that it is possible to identify the mutations solely by comparing the chromatogram of the tryptic digest with the profile obtained by computer simulation.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0021-9673
pubmed:author
pubmed:issnType
Print
pubmed:day
22
pubmed:volume
539
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
465-73
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1991
pubmed:articleTitle
Purification and characterization of recombinant tropomyosins.
pubmed:affiliation
CRBM du CNRS, INSERM U-249, Montpellier, France.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't