I want to express a recombinant protein, but I am unable to obtain the mRNA sequence encoding this protein. Therefore, I intend to use a sequence available in a database. Thus, I would like to request the synthesis of a vector containing the gene sequence, but I am unsure whether to ask for the cloning of the entire mRNA sequence (1534 bp) or just the coding sequence (1220 bp). The CDS is directly embedded within the mRNA sequence, but there are nucleotides before and after the CDS.
Hi Victor,
the choice of using the mRNA seq or the CDS depends on the heterologous expression system you are using. I am not familiar with animal cells so cannot really give you an educated opinion about it, but I guess that if you are expressing a mammalian gene in a mammalian cells the introns will not be a problem.
On the other hand if your system of choice is Saccharomyces or E.coli then I would just use the codon optimized CDS.
Hi victo pastre
It is recommended to examine the function of each segment within the protein by bioinformatics tools and utilize only the coding sequence that is necessary for your specific purpose. For example signal seq should be removed, if you want express this protein in E. coli.
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