2011
DOI: 10.1002/bit.23219
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Utilization of non‐AUG initiation codons in a flow cytometric method for efficient selection of recombinant cell lines

Abstract: Here we describe a method that couples flow cytometric detection with the attenuated translation of a reporter protein to enable efficient selection of CHO clones producing high levels of recombinant proteins. In this system, a small cell surface reporter protein is expressed from an upstream open reading frame utilizing a non-AUG initiation (alternate start) codon. Due to the low translation initiation efficiency of this alternate start codon, the majority of translation initiation events occur at the first A… Show more

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Cited by 17 publications
(12 citation statements)
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“…ClonePix™ from Genetix [202,203] and CellCelector from Aviso [204] are two systems that have been used for selecting recombinant protein-producing colonies. Cells expressing mAb are plated in semi-solid media containing a fluorescent detection antibody against the…”
Section: Clone Selectionmentioning
confidence: 99%
“…ClonePix™ from Genetix [202,203] and CellCelector from Aviso [204] are two systems that have been used for selecting recombinant protein-producing colonies. Cells expressing mAb are plated in semi-solid media containing a fluorescent detection antibody against the…”
Section: Clone Selectionmentioning
confidence: 99%
“…Nevertheless, uAUG codons appeared to be functionally most effective in repressing CDS translation. To date, only two publications analyzing human BIRC2 and yeast GCN4 have been focusing on non-AUG uORF functions at the individual transcript level (23,24). …”
Section: Determinants Of Uorf Presence or Absencementioning
confidence: 99%
“…Therefore, it is likely that Slp1 was indeed translated from the ATA codon at reduced levels, and that the reduced amounts of Slp1 are sufficient for the vegetative cell cycle but insufficient for meiosis. Although non‐ATG initiation of translation has been reported in other organisms , this is the first such example in S. pombe .…”
Section: Resultsmentioning
confidence: 81%