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In a previous effort, optimization of the interaction between MjtRNACUATyr and the E. coli native translational machinery, such as EF-Tu, was explored in order to improve the incorporation efficiency.
We have recently reported a live imaging technique to visualize nascent proteins by coupling stimulated Raman scattering (SRS) microscopy with metabolic labeling of deuterated amino acids (D-AAs) by a cell's native translational machineries.
The SgfI site was placed one base upstream of the initiation codon, which allowed the production of recombinant proteins with the native translational initiation site or with N-terminal tags using appropriate Flexi® vectors.
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Ostreococcus lines expressing CCA1 or TOC1 from their native promoters as translational fusions to firefly luciferase were previously characterized [ 14]. pCCA1 CCA1-LUCC and pCCA1 CCA1-LUClines will be referred to andCCA1-LUC and TOC1-LUC.
The resulting pPTNST108 construct contained the 7S β-conglycinin promoter, Tobacco Etch Virus TEVV) translational enhancer, native SEB signal peptide, mutant (L45R, Y89A, Y94A) SEB open reading frame (ORF), and 35S terminator.
Specifically, we produced a Strep-tagged NiG domain in a suspension culture of CHO-K1 cells in order to preserve native folding and post-translational modifications of the bait protein.
To investigate co-translational processing of native and mutant DCD, in vitro translation using a coupled T7 RNA polymerase/rabbit reticulocyte lysate system and canine pancreatic micorsomal membranes was performed.
In addition to the pairwise connections, each residue is connected to its native state so that the translational mode or the zero eigenvalue is stabilized.
To further ensure translational credibility, a native speaker of British English (with longstanding experience as an assistant in nursing and health research studies) was involved in the translation of the system of categories and quotations into English.
Native HMGA1a undergoes various post-translational modifications [1], [2].
Moreover, post-translational modifications of native Nuc, absent in hrNuc, may affect interactions in an as of yet unknown fashion.
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