Computational analysis of MARs sequences provided noticeable insights into the structure of these sequences and led us to design an artificial MAR sequence considering the essential motifs and underlain rules. As the consensus TIS, we realized the specific context in the microalgae which associates with locating the ribosome at the correct reading frame. This study also reviews the application of various in silico tools and databases to minimize the time and efforts toward genome-wide discovery of signal peptides. Bioinformatics studies unveiled the specific sequences with the highest signal score for developing secretion systems in the microalga. Furthermore, the gene optimization criteria are reviewed to redesign the coding sequence of the genes of interest. The optimized version of the gene mainly covered the host codon preference, the less structured mRNA, and exposure of TIS. We also discuss the efficient promoter, 5′-UTR, and intron (as the intragenic factors) from the closely related species to construct the specific expression vectors. At the end, the transformation systems are discussed. The expression cassettes containing the optimized genetic elements could be delivered into the microalgae cells and conferred the resistance to herbicide Basta to the transformants for at least 90 generations.
KeywordsArtificial MAR Computational biology approach Optimized genetic elements
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