Expression cassettes

The eukaryotic expression cassette is the part of an expression vector that enables production of a protein in a eukaryotic cell. The cassette consists of a eukaryotic promoter for mRNA transcription, the gene and an mRNA termination and processing signal (Poly-A signal). 

A relatively low transformation efficiency of 103-105 colonies//rg DNA [85] and multicopy integration of expression cassettes make enzyme engineering and screening difficult. However, the first successful approaches using linear, integrative expression cassettes have been reported [86]. 

The first hurdle encountered during the development of alfalfa as a recombinant protein production system was the relative inefficiency of the available expression cassettes. A study in which a tomato proteinase inhibitor I transgene was expressed in tobacco and alfalfa under the control of the cauliflower mosaic virus (CaMV) 35S promoter showed that 3-4 times more protein accumulated in tobacco leaves compared to alfalfa leaves [5]. Despite the low efficiency of the CaMV 35S promoter in alfalfa, bio-pharmaceutical production using this system has been reported in the scientific literature. Such reports include expression of the foot and mouth disease virus antigen [6], an enzyme to improve phosphorus utilization [7] and the anti-human IgG C5-1 [8]. In this last work, the C5-1 antibody accumulated to 1% total soluble protein [8]. 

Fig. 11.1 Synthetic spidroin and spidroin-ELP plant expression cassettes.

Rice is the most important staple food crop in the world and has also emerged as the model cereal species (it is the only terrestrial plant other than Arabidopsis thaliana to benefit from a completed genome sequence, and extensive EST resources are also available). Rice has a lower annual grain yield than maize (6600 kg ha-1) and the grain has a lower protein content (8%), but like maize it is easy to transform and manipulate in the laboratory, a range of useful expression cassettes have been devel-... 

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