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Why is there a discrepancy between the actual and theoretical molecular weights?
1.Post-translational modifications such as phosphorylation and glycosylation can increase molecular weight.
2.Post-translational cleavage by the protein itself can decrease molecular weight.
3.Some proteins have several splice variants, each with a different molecular weight; these should be distinguished.
4.Related to the amino acid composition (charged vs. non-charged).
5.Formation of multimers can increase molecular weight.
What factors affect prokaryotic expression?
Factors affecting prokaryotic expression include: translation initiation site, GC content, mRNA secondary structure, codon bias, choice of plasmid vector, gene/protein size, toxicity of the foreign gene to the host, gene mutations (frameshifts), and culture/induction conditions.