Literature DB >> 26172935

Correction: A Novel Humanized GLP-1 Receptor Model Enables Both Affinity Purification and Cre-LoxP Deletion of the Receptor.

Lucy S Jun, Aaron D Showalter, Nosher Ali, Feihan Dai, Wenzhen Ma, Tamer Coskun, James V Ficorilli, Michael B Wheeler, M Dodson Michael, Kyle W Sloop.   

Abstract

Entities:  

Year:  2015        PMID: 26172935      PMCID: PMC4501787          DOI: 10.1371/journal.pone.0132875

Source DB:  PubMed          Journal:  PLoS One        ISSN: 1932-6203            Impact factor:   3.240


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The expected band size in Fig 2D and the cartoon graphic in Fig 2F are incorrect. The authors have provided a corrected version of Fig 2 here.
Fig 2

Reverse transcription for PCR validation of hGLP-1R, mGlp-1r and Glp1r lines.

(A) A schematic of the gene that is expressed in the hGLP-1R mice. (B) The protein that is produced from this gene is a fusion of the mouse signal peptide (beige residues) and the human GLP-1R protein (white residues). The blue residues are those that differ between mouse and human GLP-1R. The signaling peptide is cleaved, leaving behind a human GLP-1R protein containing the C-terminal FLAG epitope (green residues). (C) cDNA was generated from total RNA isolated from islet and lung of hGLP-1R, mGlp-1r, and Glp1r mice for RT-PCR. The 5′ P915 primer annealed in human exon 8, while the 3′ P913 primer annealed to the FLAG region, a unique site within the hGLP-1R gene. This PCR product is a 588 bp fragment only observed in the hGLP-1R mice. (D) A schematic of the gene that is expressed in the Glp-1r mice. (E) Once the splice event occurs between human exon 2 and mouse exon 2, a frame-shift mutation nullifies downstream protein expression. (F) The final protein product in Glp-1r mice is a 98-aa truncation mutant. The first 36 aa’s of the mature protein encode a fraction of the GLP-1R extracellular domain, and the remaining 40 aa’s constitute missense sequence that shows no similarity to known proteins. The RT-PCR and DNA sequence analyses of the Glp-1r gene product demonstrate the Glp-1r mouse does not code for a functional GLP-1R. (G) A schematic of the wild-type (mGlp-1r) gene. (H) Using the same primer pair, PCR products from Glp-1r (372 bp) and mGlp-1r (275 bp) mice differ in size by 97 bp. (I) The wild-type GLP-1R protein is 463 aa’s including the signaling peptide.

Reverse transcription for PCR validation of hGLP-1R, mGlp-1r and Glp1r lines.

(A) A schematic of the gene that is expressed in the hGLP-1R mice. (B) The protein that is produced from this gene is a fusion of the mouse signal peptide (beige residues) and the human GLP-1R protein (white residues). The blue residues are those that differ between mouse and human GLP-1R. The signaling peptide is cleaved, leaving behind a human GLP-1R protein containing the C-terminal FLAG epitope (green residues). (C) cDNA was generated from total RNA isolated from islet and lung of hGLP-1R, mGlp-1r, and Glp1r mice for RT-PCR. The 5′ P915 primer annealed in human exon 8, while the 3′ P913 primer annealed to the FLAG region, a unique site within the hGLP-1R gene. This PCR product is a 588 bp fragment only observed in the hGLP-1R mice. (D) A schematic of the gene that is expressed in the Glp-1r mice. (E) Once the splice event occurs between human exon 2 and mouse exon 2, a frame-shift mutation nullifies downstream protein expression. (F) The final protein product in Glp-1r mice is a 98-aa truncation mutant. The first 36 aa’s of the mature protein encode a fraction of the GLP-1R extracellular domain, and the remaining 40 aa’s constitute missense sequence that shows no similarity to known proteins. The RT-PCR and DNA sequence analyses of the Glp-1r gene product demonstrate the Glp-1r mouse does not code for a functional GLP-1R. (G) A schematic of the wild-type (mGlp-1r) gene. (H) Using the same primer pair, PCR products from Glp-1r (372 bp) and mGlp-1r (275 bp) mice differ in size by 97 bp. (I) The wild-type GLP-1R protein is 463 aa’s including the signaling peptide.
  1 in total

1.  A novel humanized GLP-1 receptor model enables both affinity purification and Cre-LoxP deletion of the receptor.

Authors:  Lucy S Jun; Aaron D Showalter; Nosher Ali; Feihan Dai; Wenzhen Ma; Tamer Coskun; James V Ficorilli; Michael B Wheeler; M Dodson Michael; Kyle W Sloop
Journal:  PLoS One       Date:  2014-04-02       Impact factor: 3.240

  1 in total

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