Journal: Botanical Studies

Article Title: Sequence analysis and protein interactions of Arabidopsis CIA2 and CIL proteins

doi: 10.1186/s40529-020-00297-z

Figure Lengend Snippet: Putative CIA2-interacting proteins

Article Snippet: To explore the biological functions of CIA2, we used a bait plasmid pGBKT7 containing fused GAL4 BD and 435-aa CIA2 fragments to screen an Y2H Arabidopsis cDNA library (Clontech Mate and Plate Library - Universal Arabidopsis) for putative CIA2-interacting proteins.

Techniques:

Journal: Botanical Studies

Article Title: Sequence analysis and protein interactions of Arabidopsis CIA2 and CIL proteins

doi: 10.1186/s40529-020-00297-z

Figure Lengend Snippet: Interactions among CIA2 and CIL fragments and various proteins, as revealed by Y2H assay. Vectors pAS2-1 and pACT2 harbor GAL4 BD and GAL4 AD, respectively. The CIA2 and CIL fragments were ligated into pAS2-1 vector. Eight full-length CIA2-interacting protein candidates were ligated into pACT2 vector. Colonies were assayed for SD/-Leu-Trp autotrophy, SD/-Leu-Trp-Trp + 20 or 50 mM 3-AT autotrophy, and β-galactosidase activity. pACT2, negative control. PC, positive control containing pVA3-1 and pTD1-1

Article Snippet: To explore the biological functions of CIA2, we used a bait plasmid pGBKT7 containing fused GAL4 BD and 435-aa CIA2 fragments to screen an Y2H Arabidopsis cDNA library (Clontech Mate and Plate Library - Universal Arabidopsis) for putative CIA2-interacting proteins.

Techniques: Y2H Assay, Plasmid Preparation, Activity Assay, Negative Control, Positive Control

Journal: Botanical Studies

Article Title: Sequence analysis and protein interactions of Arabidopsis CIA2 and CIL proteins

doi: 10.1186/s40529-020-00297-z

Figure Lengend Snippet: Domain swap analysis of CIA2 and CIL fragments, as determined by Y2H assay. The pAS2-1 and pACT2 vectors harbor GAL4 BD and GAL4 AD, respectively. The CIA2 and CIL fragments were ligated into pAS2-1 or pACT2 vectors. Colonies were assayed for SD/-Leu-Trp autotrophy, SD/-Leu-Trp-Trp + 20 or 50 mM 3-AT autotrophy, and β-galactosidase activity. pACT2, negative control. PC, positive control containing pVA3-1 and pTD1-1

Article Snippet: To explore the biological functions of CIA2, we used a bait plasmid pGBKT7 containing fused GAL4 BD and 435-aa CIA2 fragments to screen an Y2H Arabidopsis cDNA library (Clontech Mate and Plate Library - Universal Arabidopsis) for putative CIA2-interacting proteins.

Techniques: Y2H Assay, Activity Assay, Negative Control, Positive Control

Journal: Botanical Studies

Article Title: Sequence analysis and protein interactions of Arabidopsis CIA2 and CIL proteins

doi: 10.1186/s40529-020-00297-z

Figure Lengend Snippet: Interactions between CIA2/CIL and CO, as assessed by Y2H assay. Vectors pAS2-1 and pACT2 harbor GAL4 BD and GAL4 AD, respectively. The CIA2 and CIL fragments were ligated into pAS2-1 vector. The full-length CO fragment was ligated into pACT2 vector. Colonies were assayed for SD/-Leu-Trp autotrophy, SD/-Leu-Trp-Trp + 20 or 50 mM 3-AT autotrophy, and β-galactosidase activity. pACT2, negative control. PC, positive control containing pVA3-1 and pTD1-1

Article Snippet: To explore the biological functions of CIA2, we used a bait plasmid pGBKT7 containing fused GAL4 BD and 435-aa CIA2 fragments to screen an Y2H Arabidopsis cDNA library (Clontech Mate and Plate Library - Universal Arabidopsis) for putative CIA2-interacting proteins.

Techniques: Y2H Assay, Plasmid Preparation, Activity Assay, Negative Control, Positive Control

Journal: Botanical Studies

Article Title: Sequence analysis and protein interactions of Arabidopsis CIA2 and CIL proteins

doi: 10.1186/s40529-020-00297-z

Figure Lengend Snippet: Sequence alignment (using MEGA-X software) of a conserved region of 10 Arabidopsis NF-YA proteins and 23 CIA2/CIL homologous proteins. Numbers in brackets at the end of each sequence indicate the corresponding aa number of each protein. Numbers above the alignment represent amino acid position (in the alignment). Black and gray backgrounds represent conserved or similar residues, respectively. Residues marked with an asterisk are important in yeast NF-YA protein. The YA1/linker/YA2 subdomain region is demarcated based on Romier et al. . The location of the NLS sequence in the CC1 motifs of homologous proteins is shown below the alignment

Article Snippet: To explore the biological functions of CIA2, we used a bait plasmid pGBKT7 containing fused GAL4 BD and 435-aa CIA2 fragments to screen an Y2H Arabidopsis cDNA library (Clontech Mate and Plate Library - Universal Arabidopsis) for putative CIA2-interacting proteins.

Techniques: Sequencing, Software

Journal: Journal of Biological Chemistry

Article Title: Transcriptional Activators and Repressors for the Neuron-specific Expression of a Metabotropic Glutamate Receptor

doi: 10.1074/jbc.m700149200

Figure Lengend Snippet: FIGURE 1. Characterization of Grm1 TISs, genomic structure, and analysis of alternative first exon expression. A, annealed sequences of human and mouse Grm1 exon Ia, as obtained by 5-RACE and public sequence data bases. The digits associated with nucleotides indicate the number of 5-RACE clones starting at that position. The height of histograms is proportional to the number of clones. The start positions of known cDNAs or expressed sequence tags are indicated by their accession number or reference. Splice donors are indicated in boldface type. B, RNase protection assay. Multiple protected bands are generated by probes encoding exon Ia and II, whereas a single band is generated by the control -actin probe (filled arrowheads, lanes 1, 3, and 5). Undigested probes are shown (open arrowheads). A high molecular weight self-annealing product (asterisk, lane 2) was produced by the undigested exon Ia probe, which however did not produce nonspecific bands due to self-protection (lane 7). C, complete genomic structure of mouse and human Grm1, based on 5-RACE data and public genomic, cDNA, and expressed sequence tag sequences. Alignments were performed by BLAST 2.0. Protein coding regions and 5- or 3-untrans- lated regions are indicated by open and gray boxes, respectively. Constitutive and alternative splicing are indicated by filled and dashed lines, respectively. The size of exons is given in bp and in kbp (in italic type) for introns. D, regional distribution of Grm1 transcripts generated from alternative first exons. The experiments shown are representative of at least four replicates. OB, olfactory bulb; CTX, cortex; HIP, hippocampus; AMY, amygdala; STR, striatum; CRB, cerebellum.

Article Snippet: 5 -Rapid Amplification of cDNA Ends (RACE)—The transcription initiation sites (TISs) of the human andmousemGlu1 gene were determined by 5 -RACE using whole mouse brain and human hippocampal and cerebellar Marathon-Ready cDNA (Clontech).

Techniques: Expressing, Sequencing, Clone Assay, Rnase Protection Assay, Generated, Control, High Molecular Weight, Produced, Alternative Splicing