Protein expression library

Young, R. A., and Davis, R. W., 1983. Efficient isolation of genes using and-body probes. Proceedings of the National Academy of Sciences U.S.A. 80 1194-1198. Using andbodies to screen protein expression libraries to isolate the structural gene for a specific protein. 

Harrington JJ et al. Creation of Genome-Wide Protein Expression Libraries Using Random Activation of Gene Expression. Nature Biotechnol 2001 19 440-445. 

Biochips can be used for either measuring differential expression between two populations or for testing for the presence of a DNA sequence (resequencing). Protein chips have been applied in expression profiling and antibody detection, binding specificities of a protein expression library and protein-protein interactions. 

Expression vectors are engineered so that any cloned insert can be transcribed into RNA, and, in many instances, even translated into protein. cDNA expression libraries can be constructed in specially designed vectors derived from either plasmids or bacteriophage A. Proteins encoded by the various cDNA clones within such expression libraries can be synthesized in the host cells, and if suitable assays are available to identify a particular protein, its corresponding cDNA clone can be identified and isolated. Expression vectors designed for RNA expression or protein expression, or both, are available. 

Santini, C., Brennan, D., Mennuni, C., Hoess, R. H., Nicosia, A., Cortese, R., and Luzzago, A. (1998). Efficient display of an HCV cDNA expression library as C-terminal fusion to the capsid protein D ofbacteriophage lambda. J. Mol. Biol. 282, 125-135. 

Membrane extracts from adult H. contortus were enriched 24-fold for cysteine protease activity by passage over a Thiol-Sepharose affinity column and the proteins obtained (abbreviated as TSBP) were clearly localized to the microvillar surface of the intestinal cells (Knox et al., 1995,1999). TSBP comprised a prominent 60 kDa protein and several minor bands between 35 and 45 kDa and 97 to 120 kDa (Fig. 13.2). Protease activity at 38, 52 and 70 kDa was attributable to cysteine proteases and at 70 and 88 kDa to serine/metalloproteases, as judged by inhibition analyses. Lectin-binding studies showed that most of the TSBPs were glycosylated. Expression library... 

Bussow K et al. A human cDNA library for high-throughput protein expression screening. Genomics 2000 65 1—8. 

Multiple lines of evidence suggest that SPF, a single protein within the 20-25 kDa pheromone fraction, is responsible for female behavioural response. First, this major protein component within the D. ocoee fraction was genetically very similar to the precursor of sodefrin (Palmer, Watts, Houck, Picard and Arnold 2007), and sodefrin is a known reproductive pheromone in newts (Kikuyama, Toyoda, Ohmiya, Matsuda, Tanaka and Hayashi 1995 Kikuyama and Toyoda 1999). Second, a separate study showed that the cDNA library of proteins expressed in male D. ocoee mental glands contained a high proportion (25%) of... 

Once the recombinant expression vectors containing the cDNA inserts are produced, they are used to transform bacteria (or other host cells) and produce cDNA (expression) libraries. Expression libraries may be used to produce recombinant proteins that in some cases have significant advantages over isolating them from natural sources ... 

Antibody coverage of the human proteome is estimated to be about 5 to 10% of all human proteins and isoforms (Valle and Jendoubi, 2003). A major bottleneck in the use of protein expression arrays is the lack of such a comprehensive set of these capture agents (Hanash, 2003). Since an equivalent of the polymerase chain reaction (PCR) process for mass amplification of low abxmdant proteins does not exist, the remaining library of proteome capture ligands will need to be generated by other means such as recombinant protein expression systems (Cahill, 2001). 

The live cell microarray may eventually displace the use of protein expression microarrays for identifying drug targets, provided that more extensive libraries of full-length cDNAs needed to express the complete... 

This pathway, which is driven by the presence of free Gfly, was modified to allow for screening of mammalian cDNA expression libraries to isolate those cDNAs that activated transcription of a pheromone-responsive promoter downstream of activated G-protein. Modifications included eliminating the pheromone receptor, replacing the endogenous yeast Ga... 

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