Mostafa R Zaher, Dalia M El-Husseini, Mohamed H El-Husseiny, Azza M El Amir, Naglaa M Hagag, Reham H Tammam
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Design and application of expression constructs for FMDV serotype O structural proteins.
Design and validate flexible constructs for recombinant expression of FMDV serotype O structural proteins of the circulating topotypes using newly designed degenerate primers. The designed degenerate primers targeting diverse topotypes enabled the successful amplification of VP0, VP1, and VP3 genes. Integration of the essential transcriptional and translational regulatory elements including T7 promoter, leader g10 sequence, and T7 terminator, as well as ribosome binding site (RBS), start and stop codons, respectively via overlap extension PCR empowered efficient expression of these proteins in E. coli. Cloned constructs expressed the target proteins of expected molecular weights: VP0 (34 kDa), VP1 (24 kDa), and VP3 (22 kDa). SDS-PAGE and Western blotting confirmed high protein yield and purity. This platform demonstrated adaptability for diagnostic and vaccine development applications. The workflow offers a robust tool for producing FMDV structural proteins concerning the circulating strains attempting to improve control measures including diagnosis and vaccinations.
期刊介绍：
Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them.
All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included.
Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields.
The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories.
Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.
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