In eukaryotes, the localization of small ribosomal subunits to mRNA transcripts requires the translation of Kozak elements at the starting site. The sequence of Kozak elements affects the translation efficiency of protein synthesis. However, whether the upstream nucleotide of Kozak sequence affects the expression of recombinant proteins in Chinese hamster ovary (CHO) cells remains unclear. In order to find the optimal sequence to enhance recombinant proteins expression in CHO cells, -10 to +4 sequences around ATG in 100 CHO genes were compared, and the extended Kozak elements with different translation intensities were constructed. Using the classic Kozak element as control, the effects of optimized extended Kozak elements on the secreted alkaline phosphatase (SEAP) and human serum albumin (HSA) gene were studied. The results showed that the optimized extended Kozak sequence can enhance the stable expression level of recombinant proteins in CHO cells. Furthermore, it was found that the increased expression level of the recombinant protein was not related with higher transcription level. In summary, optimizing extended Kozak elements can enhance the expression of recombinant proteins in CHO cells, which contributes to the construction of an efficient expression system for CHO cells.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.jbiotec.2024.06.020 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
NaP-TRAP reveals the regulatory grammar in 5'UTR-mediated translation regulation during zebrafish development.
Nat Commun
December 2024
Department of Genetics, Yale University, Yale School of Medicine, New Haven, 06510, CT, USA.
The cis-regulatory elements encoded in an mRNA determine its stability and translational output. While there has been a considerable effort to understand the factors driving mRNA stability, the regulatory frameworks governing translational control remain more elusive. We have developed a novel massively parallel reporter assay (MPRA) to measure mRNA translation, named Nascent Peptide Translating Ribosome Affinity Purification (NaP-TRAP).
View Article and Find Full Text PDFKv3.3 Expression Enhanced by a Novel Variant in the Kozak Sequence of .
Int J Mol Sci
November 2024
Institute of Human Genetics, Department of Medicine, Justus Liebig University Giessen, 35390 Giessen, Germany.
Pathogenic variants in , which encodes the voltage-gated potassium channel Kv3.3, are associated with spinocerebellar ataxia type 13. SCA13 is a neurodegenerative disease characterized by ataxia, dysarthria and oculomotor dysfunction, often in combination with other signs and symptoms such as cognitive impairment.
View Article and Find Full Text PDFA Study on the Compressive Behavior of Additively Manufactured AlSi10Mg Lattice Structures.
Materials (Basel)
October 2024
Faculty of Mechanical Engineering, University of Maribor, Smetanova 17, 2000 Maribor, Slovenia.
The mechanical behavior of the metallic components fabricated by additive manufacturing (AM) technologies can be influenced by adjustments in their microstructure or by using specially engineered geometries. Manipulating the topological features of the component, such as incorporating unit cells, enables the production of lighter metamaterials, such as lattice structures. This study investigates the mechanical behavior of lattice structures created from AlSi10Mg, which were produced using the laser beam powder bed fusion (LB-PBF) process.
View Article and Find Full Text PDFOptimization of a novel expression system for recombinant protein production in CHO cells.
Sci Rep
October 2024
International Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, No. 601 Jinsui Road, Xinxiang, 453003, Henan, China.
Article Synopsis
- Chinese hamster ovary (CHO) cells are commonly used for producing recombinant proteins, but their expression levels can be low.
- This study demonstrates that optimizing the expression vector by incorporating regulatory elements like Kozak and Leader enhances protein expression significantly.
- Additionally, modifying the CHO cell line by knocking out the Apaf1 gene using CRISPR/Cas9 technology further boosts protein production, paving the way for large-scale recombinant protein production.
Identification of the Cohesive Parameters for Modelling of Bonded Joints between Flat Composite Adherends with Thick Layer of Adhesive.
Materials (Basel)
October 2024
Faculty of Mechanical Engineering, University of West Bohemia in Pilsen, Univerzitni 2732/8, 301 00 Plzen, Czech Republic.
The failure of bonded composite materials is accompanied by specific failure modes. These are specifically Mode I, Mode II, Mode III, and their combination (so-called mixed mode). These modes depend on the direction and type of loading.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!