摘要：【目的】以Split-GFP 双荧光互补技术检测鸡肌肉生长抑制素基因（MSTN ）RNA 干涉（RNAi ）效果，并与其 他常用检测方法比较，以验证Split-GFP 双分子荧光互补技术在RNAi 效果评估中的有效性和可行性。【方法】将合成的3个shRNA 慢病毒载体（shRNA-a 、shRNA-b 和shRNA-c ）分别转染稳定表达GFP11-MSTN ...
3. Construction of a bimolecular split GFP system on mammanlian cell membrane. We obtained pDisplay-gfp1-10 recombinant plasmid with a mammalian expression vector 硕士学位论文 IV pDisplayTM, that allows GFP1-10 to appear on the extracellular side of cell membrane, Then, GFP1-10, displa...
d) GFP fluorescence intensity measured on living HEK293 cells 24h post-transfection with a single plasmid expressing either WT GFP, control (ctrl) GFP with a six-residue footprint (checkered bars), or a dual plasmid expressing split intein/GFP. The protein ligation efficiency of each split ...
B) Yeast cells expressing Mdm34-RFP from the multi-copy plasmid (pMY33) (A) or Mdm12-mScarlet from the chromosome (B) were transformed with plasmids coding for the indicated split-GFP proteins and imaged by confocal fluorescent microscopy. Maximum projection images reconstituted from z-...
As the final step towards a split GFP-based in vivo protein detection assay, target and detector proteins should be co-produced inB. subtilis. To analyze if the co-expression allows a comparative analysis of GUS11 accumulation inside live cells, a two-plasmid system was employed consisting of...
Switch fusions were transiently expressed using the FreeStyle HEK293 expression system. Briefly, transfection was initiated when the cell density of FreeStyle 293 suspension cells reached 3 × 106/mL. Subsequently, 100 μg of the expression vector plasmid was diluted in Opti-MEM (Gibco, 31...
To compare the activity of our Split-GFP-Cre system with these two systems, the components of each system were subcloned into the pMSCV-IRES-Thy1.1 expression vector. The expression level of Split-Cre was divided into high, middle, and low according to the strength of Thy1.1 expression, and...
Plasmids were transformed into DH5α electrocompetent cells for cloning and plasmid purification (Qiagen Plasmid Midi Kit) for sequencing and a cell-free reaction. Whole-Cell Protein Expression and Fluorescent Measurement The pETBlue1 plasmids containing the 1–10 and the 11th segment genes and full-...
a Cartoon of targeted split-GFP chimeras are able to complement at the OMM, within the IMS and in the mitochondrial matrix. Sub-mitochondrial localization of wild type and mutant α-syn was analyzed in HeLa b and SHSY5Y c cells by the co-expression of the OMM, IMS and mitochondrial matrix...
A plasmid encoding GFP was co-transfected as an internal transfection control. The experiments were conducted twice with similar results. The white, orange, and blue arrowheads mark PIGS-BE, Split-BE-N, and Split-BE-C, respectively. Rubisco staining indicates equal protein loading. Source data ...