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ProteoLink™ In Vitro Expression Cloning System |
 Get
a one-step direct link between gene sequences and protein functions.
ProteoLink™ System offers a one-step transcription/translation
process to identify relationships between specific cDNA sequences
and their encoded protein functions.
Promega’s ProteoLink™ In Vitro Expression Cloning System
(IVEC) is a systematic and broadly applicable tool that can be used
to directly correlate genes with proteins. The simple in vitro
method eliminates theoretical or indirect approaches to establish
gene-protein relationships.
How the ProteoLink™ System works
The system is a plasmid-based human adult brain cDNA library that is
expressed using Promega’s TNT®
Quick Coupled Transcription/Translation System (Figure 1).
Small plasmid pools are expressed in vitro circumventing the
traditional and laborious process of protein purification, peptide
sequencing and cDNA isolation to identify the genes that encode
specific proteins. 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.
Because the cDNA is cloned into a vector containing the appropriate
promoters, the identified cDNAs can be expressed in mammalian cells
to confirm the in vitro screening results in a physiological system.
Compare the ProteoLink™ System to
other approaches
To understand how genomes direct cellular function, investigators
need to identify relationships between specific DNA sequences and
their encoded protein functions. The identification of genes that
encode a particular protein of interest requires both biochemical
and genetic approaches. Traditional biochemical methods involve
purifying proteins based on a particular characteristic or function.
The genes that encode the proteins are isolated through multiple
steps that include peptide sequencing and complementary DNA (cDNA)
isolation based on predicted sequences. Other methods are also used
to analyze proteins and are used indirectly to correlate proteins to
specific cDNA sequences. Each has its own limitations for providing
a direct correlation between a specific gene and a specific protein.
Limitations of traditional approaches
for studying gene:protein relationships
2-D gel analysis provides no direct link between gene sequence and
protein function
Bioinformatics theoretical, not based on experimental data
Mass spectrometry provides no direct link between gene sequence and
protein function
Yeast Two-Hybrid Screening only defines protein:protein
interactions, does not detect cellular roles of proteins
Cell-based expression biochemical assays can be difficult, and
variability of cell culture may alter results
ProteoLink™System:
The Complete Answer
Unlike the traditional approaches outlined above, the ProteoLink™
System is designed to supply accurate data with maximum convenience,
quality and flexibility. The ProteoLink™ System...
... is Convenient because it is provided in a 96 well format.
... provides a Quality average cDNA insert size of 2.7kb.
... is a Flexible Assay for activity, substrate or interaction
screens. Extracts can be added for supplemental activity.
... is Robust. Promega’s TNT®
Quick Coupled Transcription/Translation System is used for protein
expression.
... is Fast: One-step transcription/translation process; no
secondary protein purification necessary, or cell transfections.
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