Journal: Science Advances

Article Title: Shape-morphing living composites

doi: 10.1126/sciadv.aax8582

Figure Lengend Snippet: ( A ) Schematic of a light sensitive yeast two-hybrid. Blue light induces expression of HIS3 and lacZ reporters by inducing conformational changes in CRY2 to favor interaction with CIB1. Reporter genes are transcribed by recruitment of the Gal4 activation domain (AD). ( B ) β-Galactosidase assays of an auxotrophic strain lacking CIB1 (negative control), a strain not auxotrophic for l -histidine in the dark (positive control), and the auxotrophic strain depicted in (A) (experimental). ( C ) Schematic of a living composite irradiated with blue light in growth medium lacking l -histidine. ( D ) Volume change of living composites with experimental yeast irradiated with blue light or kept in the dark. Scale bar, 5 mm. ( E ) Volume change of living composites with each yeast strain when exposed to blue light or kept in the dark. ( F ) Patterned photoresponsive living composite with the experimental yeast strain in medium lacking l -histidine where blue light is first targeted on the left side and then the right side. Scale bar, 10 mm. Each data point represents the mean ( n = 3), and error bars represent SD.

Article Snippet: L40 yeast were transformed with experimental constructs CRY2 LexA DNA binding fusion in the expression vector pDBTrp (pDBTrp-LexABD-CRY2FL) (plasmid no. 78210, Addgene) and a separate CIB1 Gal4 activation domain fusion vector pGADT7 (pGal4AD-CIB1) (plasmid no. 28245, Addgene). pDBTrp-LexABD-CRY2FL, along with the pGADT7 empty vector, was used as negative control.

Techniques: Expressing, Activation Assay, Negative Control, Positive Control, Irradiation

Journal: PLoS ONE

Article Title: Co-Interactive DNA-Binding between a Novel, Immunophilin-Like Shrimp Protein and VP15 Nucleocapsid Protein of White Spot Syndrome Virus

doi: 10.1371/journal.pone.0025420

Figure Lengend Snippet: ( A ) Photographic film of immunoblot detection of BD-VP15 fusion protein with anti c-Myc antibody, confirming its expression in transformed Y187 compared to un-transformed Y187 (-ve). ( B ) Yeast two-hybrid screen results indicating the presence of BD and AD-plasmids in transformant cells growing on an SD/-Leu/-Trp plate. Putative interaction of BD-VP15 and AD-PmFKBP46c is indicated by growth and blue color on selective medium. The positive control comprised interaction between murine p53 bait fusion and SV40 prey fusion (BD Biosciences). The negative controls comprised yeast cells containing BD-VP15 and empty pGADT7 vector or empty pGBKT7 vector and AD-PmFKBP46c.

Article Snippet: The empty vectors pGADT7 or pGBKT7 were used as negative controls and interaction between murine p53 bait fusion and SV40 prey fusion (BD Biosciences) served as a positive control.

Techniques: Western Blot, Expressing, Transformation Assay, Two Hybrid Screening, Positive Control, Plasmid Preparation

Journal: International Journal of Molecular Sciences

Article Title: Cloning and Functional Analysis of BcMYB101 Gene Involved in Leaf Development in Pak Choi ( Brassica rapa ssp. Chinensis )

doi: 10.3390/ijms21082750

Figure Lengend Snippet: Expression profiles of BcMYB101 in pak choi. ( A ) The transcript abundance of BcMYB101 at seedling, rosette, flowering, and podding stages. ANOVA was selected for statistical analysis. Letters (a, b, c, d, e) represent the difference of expression level of BcMYB101 gene in diverse tissues. ( B ) The expression analysis of BcMYB101 under gibberellin (GA3) and abscisic acid (ABA) treatments. Sixth-leaf-stage pak choi seedling were exposed to GA3 (100 μM) and ABA (100 μM) treatments over a consecutive time course (0, 1, 2, 3, 4, 6, 8, 12, 24, 36, and 48 h). The vertical and horizontal axes represent the relative expression level and time, respectively. The transcript level of BcMYB101 at 0 h was used as a control (expression value = 1). A t -test (two-tail) was selected for statistical analysis; * 0.01 < p < 0.05, ** p < 0.01.

Article Snippet: The BD- BcMYB101 -FL construct was co-transformed with an empty pGADT7 vector (AD) into yeast strain Y2H Gold, and the concrete operation scheme of yeast transformation was operated according to the instruction manual (Clontech, Mountain View, CA, USA).

Techniques: Expressing

Journal: International Journal of Molecular Sciences

Article Title: Cloning and Functional Analysis of BcMYB101 Gene Involved in Leaf Development in Pak Choi ( Brassica rapa ssp. Chinensis )

doi: 10.3390/ijms21082750

Figure Lengend Snippet: Subcellular localization of BcMYB101 . ( A ) The construct of 35S:GFP (green fluorescent protein) and 35S: BcMYB101 -GFP fusion protein. ( B ) The panels from left to right correspond to the mCherry (nuclear marker), bright-field, fluorescence, and merged fluorescence images of 35S:GFP and 35S: BcMYB101 -GFP fusion protein. Scale bars = 50 µm.

Article Snippet: The BD- BcMYB101 -FL construct was co-transformed with an empty pGADT7 vector (AD) into yeast strain Y2H Gold, and the concrete operation scheme of yeast transformation was operated according to the instruction manual (Clontech, Mountain View, CA, USA).

Techniques: Construct, Marker, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Cloning and Functional Analysis of BcMYB101 Gene Involved in Leaf Development in Pak Choi ( Brassica rapa ssp. Chinensis )

doi: 10.3390/ijms21082750

Figure Lengend Snippet: Phenotype of BcMYB101 overexpression in Arabidopsis . ( A ) Leaf number difference of wild-type (WT) and transgenic plants overexpressing BcMYB101 . Scale bar = 2 cm. ( B ) Rosette leaf number at bolting in the WT and BcMYB101 -overexpressed lines OE2, OE5, and OE6. Error bars represent the standard deviation of the mean number of 20 plants for each line. ANOVA was used for statistical analysis. Letters (a, b, c) indicate the difference of leaf number in WT and BcMYB101 -overexpressed plants. ( C ) Phenotype of downward-curling cauline leaves and higher leaf number in transgenic lines; the top view of the leaf is displayed at the bottom. Scale bar = 5 cm.

Article Snippet: The BD- BcMYB101 -FL construct was co-transformed with an empty pGADT7 vector (AD) into yeast strain Y2H Gold, and the concrete operation scheme of yeast transformation was operated according to the instruction manual (Clontech, Mountain View, CA, USA).

Techniques: Over Expression, Transgenic Assay, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Cloning and Functional Analysis of BcMYB101 Gene Involved in Leaf Development in Pak Choi ( Brassica rapa ssp. Chinensis )

doi: 10.3390/ijms21082750

Figure Lengend Snippet: Virus-induced BcMYB101 silencing in pak choi. ( A ) Expression levels of BcMYB101 in gene silencing plants. Error bars represent the standard deviation of the mean number of three replicates. ANOVA was used for statistical analysis. Letters (a, b, c, d) represent the difference of expression level of BcMYB101 gene in PTY and BcMYB101 -silencing plants. ( B ) Upward-curling leaf phenotype in pak choi plants with BcMYB101 silencing. Scale bar = 5 cm.

Article Snippet: The BD- BcMYB101 -FL construct was co-transformed with an empty pGADT7 vector (AD) into yeast strain Y2H Gold, and the concrete operation scheme of yeast transformation was operated according to the instruction manual (Clontech, Mountain View, CA, USA).

Techniques: Expressing, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Cloning and Functional Analysis of BcMYB101 Gene Involved in Leaf Development in Pak Choi ( Brassica rapa ssp. Chinensis )

doi: 10.3390/ijms21082750

Figure Lengend Snippet: Trans-activation activity analysis of BcMYB101. Transcription activity assay of BcMYB101 in yeast. BcMYB101 -FL, BcMYB101 -N, BcMYB101 -M, and BcMYB101 -C represent the full-length protein, as well as the regions encoding the N-terminal, middle, and C-terminal regions of BcMYB101 , respectively, which were inserted into the pGBKT7 vector. The number of amino-acid residues is shown above the panel.

Article Snippet: The BD- BcMYB101 -FL construct was co-transformed with an empty pGADT7 vector (AD) into yeast strain Y2H Gold, and the concrete operation scheme of yeast transformation was operated according to the instruction manual (Clontech, Mountain View, CA, USA).

Techniques: Activation Assay, Activity Assay, Plasmid Preparation

Journal: International Journal of Molecular Sciences

Article Title: Cloning and Functional Analysis of BcMYB101 Gene Involved in Leaf Development in Pak Choi ( Brassica rapa ssp. Chinensis )

doi: 10.3390/ijms21082750

Figure Lengend Snippet: Verification of candidate proteins interacting with BcMYB101. pGADT7 + pGBKT7 and pGADT7-T + pGBKT7-53 were used as the negative and positive control. The blue triangle represents the concentration of yeast solution from high to low (10 0 –10 −3 ).

Article Snippet: The BD- BcMYB101 -FL construct was co-transformed with an empty pGADT7 vector (AD) into yeast strain Y2H Gold, and the concrete operation scheme of yeast transformation was operated according to the instruction manual (Clontech, Mountain View, CA, USA).

Techniques: Positive Control, Concentration Assay