[Optimizing the expression of Mx gene in Escherichia coli based on rare codon and mRNA structure].

Yi Chuan

National Center for Molecular Genetics and Breeding of Animal, Institute of BeiJing Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China.

Published: January 2009


Category Ranking

98%

Total Visits

921

Avg Visit Duration

2 minutes

Citations

20

Article Abstract

Rare codon and mRNA secondary structure of translation initiation region were analyzed. Four bioengineered bacterium BL21(DE3)/pET-Mx, Rosseta (DE3)/pET-Mx, BL21(DE3)/pGEX-Mx, and Rosseta (DE3)/pGEX-Mx were obtained. Through optimizing the rare codon and mRNA secondary structure of translation initiation region, Mx protein was expressed in Rosseta (DE3)/pET-Mx, and Rosseta (DE3)/pGEX-Mx. The specified product of 75 kDa was detected by means of Western blotting analysis. The result showed that Rosetta (DE3) strain expressing some rare codon used in our experiment can obtain Mx protein expression, and the lower energy of mRNA structure can improve the expression level of Mx protein. This is the first report on the expression of the full open reading frame of Mx gene.

Download full-text PDF

Source
http://dx.doi.org/10.3724/sp.j.1005.2009.00075DOI Listing

Publication Analysis

Top Keywords

rare codon
16
codon mrna
12
mrna secondary
8
secondary structure
8
structure translation
8
translation initiation
8
initiation region
8
rosseta de3/pet-mx
8
rosseta de3/pgex-mx
8
[optimizing expression
4

Similar Publications

Introduction: Metastatic colorectal cancer (mCRC) exhibits significant heterogeneity in molecular profiles, influencing treatment response and patient outcomes. Mutations in v-raf murine sarcoma viral oncogene homolog B1 () and rat sarcoma () family genes are commonly observed in mCRC. Though originally thought to be mutually exclusive, recent data have shown that patients may present with concomitant and mutations, posing unique challenges and implications for clinical management.

View Article and Find Full Text PDF

Rationale: Weaver syndrome is a rare congenital overgrowth disorder characterized by a wide spectrum of clinical manifestations that often overlap with other overgrowth syndromes. It is primarily caused by pathogenic variants in the Enhancer of Zeste Homolog 2 (EZH2) gene on chromosome 7q36.1.

View Article and Find Full Text PDF

The AUA isoleucine codon is generally rare and used with varying frequency in bacterial genomes. The tRNA responsible for decoding this trinucleotide must be modified at the wobble position by tRNA lysidine synthetase (TilS) prior to aminoacylation and accommodation at the ribosome. To test the hypothesis that TilS catalytic efficiency correlates with AUA frequency, we cloned tilS genes from bacteria with varying AUA codon usage.

View Article and Find Full Text PDF

Background: Fishes are key components of the megafauna of the deep sea, and evolutionary adaptations to deep-sea life appear to have occurred independently in at least 22 fish orders. In this context, the analysis of even more fish genomes and mitogenomes has fundamental importance, providing a valuable resource for understanding the molecular mechanisms underlying evolutionary adaptation, especially to extreme environments such as the deep sea. Here, we report the first complete mitochondrial genome of Zu cristatus (Bonelli, 1819), providing essential information on its structure and phylogeny.

View Article and Find Full Text PDF

Viruses use a range of sophisticated strategies to evade detection by cytotoxic T-lymphocytes (CTLs) within host cells. Beyond elaborating dedicated viral proteins that disrupt the MHC class I antigen-presentation machinery, some viruses possess intrinsic, cis-acting genome-encoded elements that interfere with antigen processing and display. These protein features, including G-quadruplex motifs, repetitive peptide sequences, and rare-codon usage, counterintuitively limit production of proteins critical to virus survival, particularly during latency.

View Article and Find Full Text PDF