scholarly journals LCR/MEL: A versatile system for high-level expression of heterologous proteins in erythroid cells

1992 ◽  
Vol 20 (5) ◽  
pp. 997-1003 ◽  
Author(s):  
M. Needham ◽  
C. Gooding ◽  
K. Hudson ◽  
M. Antoniou ◽  
F. Grosveld ◽  
...  
Keyword(s):  
Heterologous Proteins ◽  
Erythroid Cells ◽  
High Level Expression ◽  
High Level ◽  
In Erythroid Cells

High-Fidelity Translation of Recombinant Human Hemoglobin in Escherichia coli

1998 ◽  
Vol 64 (5) ◽  
pp. 1589-1593 ◽  
Author(s):  
Michael J. Weickert ◽  
Izydor Apostol
Keyword(s):  
Escherichia Coli ◽  
Error Rates ◽  
High Fidelity ◽  
Human Hemoglobin ◽  
Heterologous Proteins ◽  
Expression Systems ◽  
High Level Expression ◽  
E Coli ◽  
Translation Error ◽  
High Level

ABSTRACT Coexpression of di-α-globin and β-globin in Escherichia coli in the presence of exogenous heme yielded high levels of soluble, functional recombinant human hemoglobin (rHb1.1). High-level expression of rHb1.1 provides a good model for measuring mistranslation in heterologous proteins. rHb1.1 does not contain isoleucine; therefore, any isoleucine present could be attributed to mistranslation, most likely mistranslation of one or more of the 200 codons that differ from an isoleucine codon by 1 bp. Sensitive amino acid analysis of highly purified rHb1.1 typically revealed ≤0.2 mol of isoleucine per mol of hemoglobin. This corresponds to a translation error rate of ≤0.001, which is not different from typical translation error rates found for E. coli proteins. Two different expression systems that resulted in accumulation of globin proteins to levels equivalent to ∼20% of the level of E. colisoluble proteins also resulted in equivalent translational fidelity.

High level expression of heterologous proteins in methylotrophic yeastPichia pastoris

1988 ◽  
Vol 28 (4) ◽  
pp. 265-278 ◽  
Author(s):  
K. Sreekrishna ◽  
Rica H. B. Potenz ◽  
John A. Cruze ◽  
William R. McCombie ◽  
Kathryn A. Parker ◽  
...  
Keyword(s):  
Heterologous Proteins ◽  
High Level Expression ◽  
High Level

scholarly journals Calpeptin Increases the Activity of Upstream Stimulatory Factor and Induces High Level Globin Gene Expression in Erythroid Cells

2009 ◽  
Vol 284 (30) ◽  
pp. 20130-20135 ◽  
Author(s):  
I-Ju Lin ◽  
Zhuo Zhou ◽  
Valerie J. Crusselle-Davis ◽  
Babak Moghimi ◽  
Kunjal Gandhi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Globin Gene ◽  
Erythroid Cells ◽  
Upstream Stimulatory Factor ◽  
High Level ◽  
Stimulatory Factor ◽  
Globin Gene Expression ◽  
In Erythroid Cells

scholarly journals The KH-Domain Protein αCP Has a Direct Role in mRNA Stabilization Independent of Its Cognate Binding Site

2003 ◽  
Vol 23 (4) ◽  
pp. 1125-1134 ◽  
Author(s):  
Jian Kong ◽  
Xinjun Ji ◽  
Stephen A. Liebhaber
Keyword(s):  
Unique Contribution ◽  
Erythroid Cells ◽  
Binding Studies ◽  
Globin Mrna ◽  
Direct Role ◽  
Complex Assembly ◽  
Mrna Stabilization ◽  
High Level ◽  
In Erythroid Cells

ABSTRACT Previous studies suggest that high-level stability of a subset of mammalian mRNAs is linked to a C-rich motif in the 3′ untranslated region (3′UTR). High-level expression of human α-globin mRNA (hα-globin mRNA) in erythroid cells has been specifically attributed to formation of an RNA-protein complex comprised of a 3′UTR C-rich motif and an associated 39-kDa poly(C) binding protein, αCP. Documentation of this RNA-protein α-complex has been limited to in vitro binding studies, and its impact has been monitored by alterations in steady-state mRNA. Here we demonstrate that αCP is stably bound to hα-globin mRNA in vivo, that α-complex assembly on the hα-globin mRNA is restricted to the 3′UTR C-rich motif, and that α-complex assembly extends the physical half-life of hα-globin mRNA selectively in erythroid cells. Significantly, these studies also reveal that an artificially tethered αCP has the same mRNA-stabilizing activity as the native α-complex. These data demonstrate a unique contribution of the α-complex to hα-globin mRNA stability and support a model in which the sole function of the C-rich motif is to selectively tether αCP to a subset of mRNAs. Once bound, αCP appears to be fully sufficient to trigger downstream events in the stabilization pathway.

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