A premature termination of human epidermal growth factor receptor transcription in Escherichia coli.
Our success in producing an active epidermal growth factor receptor (EGFR) tyrosine kinase in Escherichia coli encouraged us to express the full-length receptor in the same host. Despite its large size, we were successful at producing the full-length EGFR protein fused to glutathione S-transferase (GST) that was detected by Western blot analysis. Moreover, we obtained a majoritarian truncated GST-EGFR form detectable by gel electrophoresis and Western blot. This truncated protein was purified and confirmed by MALDI-TOF/TOF analysis to belong to the N-terminal extracellular region of the EGFR fused to GST. Northern blot analysis showed two transcripts suggesting the occurrence of a transcriptional arrest.
Centre of Biotechnology of Sfax, P.O. Box 1177, 3018 Sfax, Tunisia.
Jihene
J
Elloumi-Mseddi
Centre of Biotechnology of Sfax, P.O. Box 1177, 3018 Sfax, Tunisia.
Karim
K
Jellali
Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Científica and Universidad Autónoma de Madrid, Arturo Duperier 4, 28029 Madrid, Spain.
Antonio
0000-0002-4200-374X
A
Villalobo
Centre of Biotechnology of Sfax, P.O. Box 1177, 3018 Sfax, Tunisia.
Sami
0000-0003-1466-5890
S
Aifa
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D018389:Codon, Nonsense; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
D018389:Codon, Nonsense; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; C512478:EGFR protein, human; D066246:ErbB Receptors