Cis/transgene optimization: systematic discovery of some key gene expression elements integrating bioinformatics and computational biology

摘要: In recombinant protein production, quantity and quality are the major challenges particularly for large scale high-throughput production systems. The present study mainly focused on computational analysis in silico systematic discovery of some key functional gene expression elements microalgae Dunaliella salina as a case which there is no or poor information this regard. Among factors, we took shot at matrix attachment regions (MARs), translation initiation sites (TIS), signal peptide (SP) sequences, optimization transformation system. Computational MARs sequences provided enough about structure these led us to design an artificial MAR sequence considering essential motifs underlain rules. As consensus TIS, revealed that A-3, G-6C-5C-4 G+1C+2G+3 arrange specific context help locating ribosome correct reading frame. Bioinformatics studies unveiled MASTRAPLLALLALLCAGSARA with highest score SP secretion A multi-criteria procedure was performed redesign coding BAR selectable marker gene. optimized version covered host codon preference, less structured mRNA exposure TIS. intragenic selected efficient promoter, 5ˈ-UTR intron from closely related species (Chlamydomonas Sp.) construct vectors. cassettes containing genetic could be delivered into cells conferred resistance transformants least 90 generations. findings indicated flanking cassette along adaptation potentially improve efficiency stability. can efficiently deployed empirical model cis/transgenes.