polyclonal rabbit anti-peptide sera  (GenScript corporation)

Journal: Journal of Experimental Botany

Article Title: Irregular green netting of eggplant fruit peel: a domestication trait controlled by SmGLK2 with potential for fruit colour diversification

doi: 10.1093/jxb/erae355

Figure Lengend Snippet: From the genomic sequence of the SmGLK2 gene to protein structure. (A) SmGLK2 annotated gene structure in 67/3, HQ-1315, and GUIQIE-1 eggplant reference genomes and SL4.0 tomato genome [5ʹ-untranslated region (UTR) in blue, eggplant annotated exons in yellow, tomato annotated exons in red, and 3ʹ-UTR in green]. (B) The S. melongena 67/3 (SRR3884608), S. incanum INC1 (SRR2289250), and S. insanum MM0686 accession (SRR8736646) transcripts aligned against the SmGLK2 gene sequence from the 67/3 v.3 eggplant reference genome and visualized in the IGV tool ( Robinson et al. , 2023 ). The candidate structural variation identified is indicated with a purple line. (C) The suggested SmGLK2 gene structural annotation as a consensus of previous information with the extra proposed exon in purple. The identified structural variation in the gene sequence is indicated with a black arrowhead and a red line. (D) Comparison of mRNA sequences for presence (FN) or absence (fn) of the fruit green netting trait. The premature stop codon downstream of the indel is indicated with an asterisk. (E) From left to right: comparison of the protein structure and sequence around the indel site for FN and fn with the nuclear localization signal (NLS) indicated in dark grey and the golden2-like transcription factor domain in green; western blot showing the difference in apparent molecular mass of the GLK2 protein from FN and fn tissues; and differences in the expression level of GLK1 and GLK2 from different tissues of S. insanum INS1. PP2AA2 gene was used as a reference gene for normalization. Columns and error bars represent means and standard deviations of three replicates. Different letters above the columns indicate statistical significance ( P <0.05) according to Student’s t -test.

Article Snippet: The anti-sera peptide-based polyclonal rabbit GLK2-antibodies were raised against the SmGLK2 N-terminal sequence (synthetic peptide VSPPLSYTNENENY, 5–18 aa; and NMKSKSKEAKKSSG, 70–83 aa) (Davids Biotechnologie GmbH, Regensburg, Germany).

Techniques: Sequencing, Comparison, Western Blot, Expressing

Journal: Journal of Experimental Botany

Article Title: Irregular green netting of eggplant fruit peel: a domestication trait controlled by SmGLK2 with potential for fruit colour diversification

doi: 10.1093/jxb/erae355

Figure Lengend Snippet: Maximum likelihood dendrogram of GLK proteins from Arabidopsis and Solanaceae species (eggplant, tomato, and pepper). In orange, GLK1-like proteins and in green, GLK2-like proteins. AT, Arabidopsis thaliana ; SMEL, Solanum melongena (eggplant); Solyc, S. lycopersicum (tomato); CA, Capsicum annuum (pepper). Scale bar refers to a 0.10 distance and gene/IDs correspond to those from Solgenomics.net.

Article Snippet: The anti-sera peptide-based polyclonal rabbit GLK2-antibodies were raised against the SmGLK2 N-terminal sequence (synthetic peptide VSPPLSYTNENENY, 5–18 aa; and NMKSKSKEAKKSSG, 70–83 aa) (Davids Biotechnologie GmbH, Regensburg, Germany).

Techniques:

Journal: bioRxiv

Article Title: The irregular fruit green netting: An eggplant domestication trait controlled by the SmGLK2 gene with implications in fruit colour diversification

doi: 10.1101/2023.06.28.546667

Figure Lengend Snippet: From the genomic sequence of the SmGLK2 gene to protein structure. (A) SmGLK2 annotated gene structure in 67/3, HQ-1315, and GUIQIE-1 eggplant reference genomes and SL4.0 tomato genome (5’-UTR in blue, eggplant annotated exons in yellow, tomato annotated exons in red, and 3’-UTR in green). (B) The S. melongena 67/3 (SRR3884608), S. incanum INC (SRR2289250), and S. insanum MM0686 accession (SRR8736646) transcripts aligned against the SmGLK2 gene sequence from the 67/3 v.3 eggplant reference genome and visualized in the IGV tool ( Robinson et al ., 2023 ). The candidate structural variation identified is indicated with a purple line. (C) The suggested SmGLK2 gene structural annotation as a consensus of previous information with the extra proposed exon in purple. The identified structural variation in the gene sequence is indicated with a black arrowhead and a red line. (D) Comparison of mRNA sequences for presence (FN) or absence (fn) of the fruit green netting trait. The premature stop codon downstream of the indel is indicated with an asterisk. (E) Comparison of the protein structure and sequence around the indel site for FN and fn. The golden-2 like transcription factor domain is indicated in green. On the right, a Western blot showing the difference in apparent molecular mass of the GLK2 protein from FN and fn tissues.

Article Snippet: The anti-sera peptide based polyclonal rabbit GLK2-antibodies were raised against the SmGLK2 N-terminal sequence (synthetic peptide VSPPLSYTNENENY, 5–18 aa; and NMKSKSKEAKKSSG, 70–83 aa) (Davids Biotechnologie GmbH, Regensburg, Germany).

Techniques: Sequencing, Comparison, Western Blot