Various techniques have been developed to screen large expressed protein libraries for proteins or protein fragments with specific binding properties, such as the yeast two-hybrid system and phage display. However, these methods have limitations since yeast two-hybrid system relies on the internal co-expression of “bait” and “prey” fusion proteins, and the phage display is limited by potential expression bias. To address these constraints, yeast surface-displayed libraries of human cDNA fragments can be utilized. Through our unparalleled yeast display platform, Creative Biolabs offers integrated service of cDNA library construction to our worldwide customers.
Yeast Display cDNA Library
Complementary DNA (cDNA) is the reverse transcript of mRNA. As lacking the large non-coding introns compared to genomic DNA, eukaryotic cDNA libraries represent the coding sequence of all temporally transcribed genes in a certain cell type. Heterologous protein fragments can be efficiently displayed at high copy levels on the Saccharomyces cerevisiae cell wall via yeast display system. Yeast display is widely used to screen large expressed protein libraries for proteins or protein fragments with specific binding properties.
Recently, yeast surface displayed cDNA libraries have been used to identify proteins which bind to various target molecules such as peptides, antibodies, and small molecules. Since yeast protein expression pathways are like those found in mammalian cells, human protein fragments displayed on the yeast cell wall are most likely to be properly folded and functional. Combined with fluorescence-activated cell sorting (FACS), yeast display cDNA libraries allow the selection of protein fragments or domains with affinity for any soluble molecule that can be fluorescently detected.
The standard procedure to construct yeast display cDNA libraries is performed in following steps:
1) Extraction of RNA from indicated samples
2) Synthesis of cDNA candidate pool
3) Generation of frameshift variants of pYD1 vector
4) Library construction
5) Transformation of library into yeast
6) Test library induction
Fig 1. The system for displaying the cDNA library on the yeast surface and the process for enriching the target clones (Bidlingmaier and Liu, 2011).
Why Choose Creative Biolabs?
Based on extensive experience and our legendary yeast display platform, Creative Biolabs is specialized in constructing high-quality and optimized custom cDNA libraries. At Creative Biolabs, our scientists have established various high-complexity cDNA libraries cloned into an efficient vector and transformed into yeast, which can significantly reduce the labor and time needed to perform a yeast two-hybrid screen since library amplification and yeast transformation have already been done. What’s more, we can selectively eliminate highly abundant transcripts from the libraries to enhance the representations of low-abundance and rare cDNAs, in order to minimize the possibility of obtaining false positives during screening.
If you are interested in learning more about Creative Biolabs’ yeast display platform, please contact us for more details. We are also proud to provide other yeast display library construction services including: