mRNA molecules contain specific signals that optimise their interaction with ribosomes; known as leader sequences
, these include the Shine-Dalgarno (SD) sequence required for canonical translation initiation in bacteria.
After assembly of the leader sequences and the constant regions, the variable regions were then introduced via the SgfI and the SbJI sites, or the AscI and SwaI sites, respectively.
For expression of selected scFv as different Ig isotypes in mammalian cells, we constructed a set of modular cassettes containing leader sequences, constant Ig regions, and restriction sites for the incorporation of scFv or separate variable regions.
After introduction of the leader sequence into the mammalian expression vector pcDNA3.
5,834,246, is entitled "Recombinant Systems for the Expression of the Cholera B-Subunit with the Aid of Foreign Promoters and/or Leader Sequences
2] terminal leader sequences (7); the leader sequences for rcTnI-6 and rcTnI-14 are MASMGS and MASMTGGQQMGRGS, respectively.
The observed differences in immunoactivity among rcTnI-0, rcTnI-6, rcTnI-14, and their complexes can be interpreted as changes in the accessibility of given epitopes of the rcTnIs resulting from the addition of leader sequences.
In conclusion, we have demonstrated for the first time that fused leader sequences affect the immunoactivity of a recombinant protein, presumably by changing its folding status.
5,629,172, entitled "Expression of Fusion Polypeptides Transported Out of the Cytoplasm Without Leader Sequences
," relates to recombinant protein production using fusion protein technology.