In vitro expression cloning system avoids the traditional and laborious processes involved in identifying gene/protein relationships by using a one step transcription/translation process
Promega's ProteoLink in vitro expression cloning system is an efficient and widely applicable method for the characterisation and biochemical screening of cDNAs and their encoded proteins.
Avoiding the traditional and laborious processes involved in identifying gene/protein relationships, this one step transcription/translation process can be completed in hours instead of days - there is no need for cell transfection or the isolation of protein from cells.
Based on Promega's TNT quick coupled transcription/translation system, proteins are expressed in vitro from small pools of a cDNA library (human adult brain).
Each protein pool is screened for a desired biochemical activity.
Positive pools are progressively subdivided until a single cDNA encoding the active protein is isolated. Convenient to use, both the TNT Lysate and cDNA library are provided in a 96-well format.
The average cDNA insert size within this library is 2.7kb.
The cDNA, once identified, can be cloned into your vector of choice containing the appropriate promoter for expression in mammalian cells - so in vitro screening results can be confirmed in a physiological system.
Highly versatile, a variety of activity, substrate or interaction screens can be performed.
The ProteoLink system can be used, for example, to rapidly identify cDNAs that express a desired functional activity, discover proteins that interact with 'bait' (eg, ligand, multimeric protein, nucleic acids), discover genes that have low sequence homology but similar function, detect modified versions of proteins using specific antibodies, identify properties of proteins with activities under unusual conditions or identify mutant proteins with novel characteristics (ie, protein evolution).
