Codon adjustment to maximise heterologous gene expression inStreptomyces lividans can lead to decreased mRNA stability and protein yield

Lammertyn, Elke; Mellaert, Lieve Van; Bijnens, AP; Joris, Bernard; Anné, Jozef

doi:10.1007/bf02174182

Cited by 11 publications

(6 citation statements)

References 27 publications

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“…Besides codon bias, RNA secondary structure is another important factor restructured by codon change and represents significant effects on heterologous gene expression and hence should be placed under great observation during codon optimization. Lower folding energies (higher stability) slow down the ribosome and decrease translation efficiency (Lammertyn et al 1996;Tuller et al 2010). On the other hand, it appears that codon bias in lower eukaryotes tends towards A+T (Sinclair and Choy, 2002).…”

Section: Fig 5 Graphs Related To Comparison Of Secreted and Intracelmentioning

confidence: 97%

Elevating the expression level of biologically active recombinant human alpha 1-antitrypsin in Pichia pastorisan alpha 1-antitrypsin in Pichia pastoris

Arjmand¹,

Lotfi²,

Shamsara³

et al. 2013

Electron. J. Biotechnol.

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Background: Human alpha 1-antitrypsin (AAT) is a potent inhibitor of multiple serine proteases, and protects tissues against their harmful effects. Individuals with reduced or abnormal production of this inhibitor need intravenous administration of exogenous protein. In this study, we employed the methylotrophic (methanol utilizing) yeast Pichia pastoris (P. pastoris) as a preferential host for efficient production and secretion of recombinant AAT. Furthermore, we examined different strategies to maximize the yield of the secreted protein.Results: Our findings revealed that optimizing the codon usage of AAT gene for P. pastoris had positive effects on the level of secreted AAT under the control of inducible alcohol oxidase 1 (AOX1) and constitutive glycerol aldehyde phosphate dehydrogenase (GAP) promoters. Compared to AOX1, the GAP promoter increased the yield of AAT by more than two fold. It was also demonstrated that the human AAT native signal sequence was more effective than the well-known yeast signal sequence, alpha mating factor (α-MF). Doubling gene dosage nearly doubled the production of AAT, though dosages exceeding this limit had negative effects on the yield. Conclusion: P. pastoris is shown to be an efficient expression system for production of recombinant and biologically active AAT. Also different strategies could be used to elevate the amount of this secretable protein.

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Section: Fig 5 Graphs Related To Comparison Of Secreted and Intracelmentioning

confidence: 97%

Elevating the expression level of biologically active recombinant human alpha 1-antitrypsin in Pichia pastorisan alpha 1-antitrypsin in Pichia pastoris

Arjmand¹,

Lotfi²,

Shamsara³

et al. 2013

Electron. J. Biotechnol.

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“…Increased resistance against infection can be achieved in two ways: by (1) biasing the anticodons of the host tRNA pool away from the parasite codon set through reduced anticodon-codon binding specificity, or (2) modifying the copy number of a tRNA anticodon required by the parasite. These strategies can be effective as the translation of highly expressed parasite genes requires large numbers of complementary anticodons in the host (Bennetzen & Hall, 1982;Kurland, 1991;Lammertyn et al, 1996). Modifying anticodons and tRNA availabilities restrict virus genomes access to host protein-synthesis machinery.…”

Section: Translational Parasitismmentioning

confidence: 99%

Red Queen Dynamics of Protein Translation

Krakauer

Jansen

2002

Journal of Theoretical Biology

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We explore adaptive theories for the diversity of translational binding based on the genetic code viewed as a primitive mechanism of resistance. Modifying the set of codons bound by tRNA anticodon molecules or changing the specificity of binding, reduces the replication rate of translational parasites such as viruses. Increased translational efficiency of the parasite requires a high degree of specificity of host tRNAs for the parasite codons. This suggests that the genetic code might serve as the first line of defense against infection. We construct a red queen theory for translational diversity: a theory in which host-translational strategiesF as defined by the degree of redundancy (a single anticodon binding many codons for a single amino acid) or degeneracy (many anticodons binding many codons for a single amino acid)Fare constantly shifting through time to evade parasitism but where neither parasite nor host gain a systematic advantage. r

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“…The presence of rare codons would be expected to have a similar effect on mRNA stability as slowing ribosomes, and in fact, most studies show that the removal of rare codons stabilizes mRNA (Comer et al, 1996; Deana et al, 1996; Gursky and Beabealashvilli, 1994; Makoff et al, 1989). However, since rare codons result in a localized retardation of ribosome movement on mRNA as compared to bulk translation slowing by antibiotics, the removal of rare codons does not always result in mRNA stabilization but can also result in decreased mRNA stability (Lammertyn et al, 1996). These seemingly conflicting experimental observations can be explained by the location of the rare codons with respect to the mRNA cleavage positions.…”

Section: Mrna Stability Influencesmentioning

confidence: 99%

Controlling Messenger RNA Stability in Bacteria: Strategies for Engineering Gene Expression

Carrier

Keasling

1997

Biotechnol. Prog.

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Recent advances in the understanding of prokaryotic gene expression have led scientists to look beyond traditional promoter control for new methods of regulating gene expression. A promising, new technique centers on controlling the stability of messenger RNA. To exploit the potential of mRNA stability for gene expression control, it is important to understand the mechanisms of prokaryotic mRNA decay as well as the cellular factors that can be used to enhance bacterial gene expression through mRNA stabilization. Factors involved in controlling prokaryotic mRNA stability such as nucleases, secondary structures, translation influences, and transcription effects are discussed and analyzed within the context of three prevailing mRNA decay theories. Several strategies for manipulating mRNA stability in genetically-engineered cells are developed from these discussions and presented as a future direction in gene expression control. In the near future, it should be possible to use these strategies to control mRNA stability in such applications as pharmaceutical protein production and metabolic pathway design.

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Codon adjustment to maximise heterologous gene expression inStreptomyces lividans can lead to decreased mRNA stability and protein yield

Cited by 11 publications

References 27 publications

Elevating the expression level of biologically active recombinant human alpha 1-antitrypsin in Pichia pastorisan alpha 1-antitrypsin in Pichia pastoris

Elevating the expression level of biologically active recombinant human alpha 1-antitrypsin in Pichia pastorisan alpha 1-antitrypsin in Pichia pastoris

Red Queen Dynamics of Protein Translation

Controlling Messenger RNA Stability in Bacteria: Strategies for Engineering Gene Expression

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