The antigen-binding capacity of nanobodies is similar to conventional antibodies for three reasons. First, the complementarity-determining region 3 (CDR3) of nanobodies is similar or even longer than that of human VH domain (variable domain of heavy immunoglobulin chain). Second, nanobodies can form finger-like structures to recognize cavities or hidden epitopes that are not available to mAbs.
The most common yeast display system employs fusion of the protein of interest to the C-terminus of the a-agglutinin mating protein Aga2p subunit, a technology pioneered by Boder and Wittrup. The yeast surface display construct designed for this system includes two epitope tags: a hemagglutinin (HA) tag between Aga2p and the N-terminus of the protein of interest, and a C-terminal c-myc tag.
Saccharomyces cerevisiae belongs to eukaryote, which is closer to the expression system of plants and animals. This expression system includes the following processes: glycosylation, disulfide bond formation, and Post-translational modification of protein folding.
This kit is used to amplify fragments from the yeast genome as well as transformed plasmids. The component contains a visualization green dye, which can directly perform polyacrylamide gel electrophoresis and agarose gel electrophoresis after PCR. The PCR product has A- tailing at the 3' end, which is suitable for TA cloning.
Yeast one-hybrid system is a technique developed from yeast two-hybrid to study DNA-protein interactions, and is widely used to study the regulation of gene expression in eukaryotic cells, such as identifying DNA binding sites to discover potential binding protein genes, analyzing DNA binding structural domain information, etc.
TF-centered YIH (Yeast One-Hybrid Technology), which recognizes transcription factors as its recognition elements, can accurately and efficiently identify transcription factor recognition elements, and has a wide range of applications in the study of protein-DNA element interactions.
One of the most important branches of genetic engineering is the expression of recombinant proteins using biological expression systems. Nowadays, different expression systems are used for the production of recombinant proteins including bacteria, yeasts, molds, mammals, plants, and insects.
Currently, the most widely used method for prediction of signal peptide from amino acid sequences is to use software SignalP (Petersen et al., 2011). Yeast signal sequence trap (YSST) is a method that can be used to evaluate the function of predicted signal peptides. This method utilizes the yeast Saccharomyces cerevisiae YTK12 strain and pSUC2 vector in which the pSUC2 vector with fused predicted signal peptide is transformed into yeast.
The yeast three-hybrid system is mainly used to study more complex macromolecular interactions including three components, providing a new method for the study of protein-protein-protein, protein-RNA-protein, and protein-small molecule-protein interactions.
The yeast two-hybrid system is a research method for identifying and detecting protein interactions in living cells, which is performed in the eukaryotic model organism yeast, and is now used in several research fields because of its high sensitivity and wide applicability.