Finally, rDNA arrays were recently shown to influence gene expression at ectopic positions in Drosophila [ 77], and locus size could potentially serve as an epigenetic regulator harmonizing the expression of subgenomes.
We propose that progressive RNA-mediated transcriptional gene silencing plays an important role also in natural situations by harmonizing gene expression during allopolyploidy and stress.
Three hours after IPTG induction, expression levels of codon harmonized MSP142 (3D7) from pET AT) MSP142 3D7.2 were much higher than from the native sequence (Fig. 5A, WT and CH, respectively, lanes 3), and product yield increased four-fold, from 0.8 mg/g wet cells for the native sequence to 3.0 mg/g wet cells from the codon harmonized sequence (Fig 5A, Table 1).
In this current work we explain the algorithm in detail, and apply it to the expression of the codon harmonized gene sequences for three additional malaria vaccine candidates, namely MSP142 (FVO), MSP142 (3D7) and MSP142 (Camp).
Expression of the codon harmonized LSA-NRCH malaria vaccine candidate, which has been described elsewhere [26], has proven to be the best approach, to date, for production of this protein in E. coli.
Expression levels for codon harmonized MSP142 Catp.2, at 12.0 mg/g wet cells (Fig 5B, Table 1) were very high; we were not able to express wild type MSP142 (Camp) in E. coli (Table 1).
First, we generated a recombinant PfCelTOS in the E. coli expression system using the codon harmonized gene sequence from P. falciparum.
Before associating expression and phenotype data, we downloaded and harmonized data sets from several different data repositories.
Expression levels for soluble protein from the codon harmonized MSP142 3D7.2 and MSP142 Camp.2 genes equaled the levels produced for FMP010.
Internal and external information were harmonized between the Walloon and MACE evaluations by adjusting scales and mean differences towards the original expression of the trait in the Walloon genetic evaluations.
