Efficient and stable expression of recombinant fibronectin polypeptides

We describe retroviral expression vectors containing cDNAs encoding part of fibronectin preceded by the signal and 'pro' sequences of parathyroid hormone. The recombinant retroviruses were used to generate NIH 3T3 cell lines stably producing functionally active fragments of fibronectin. The recombinant fibronectins (deminectins) are processed and secreted by the cells and form disulfide-bonded dimers with themselves and with endogenous fibronectin subunits. The fibronectin-deminectin heterodimers are incorporated into the extracellular matrix. We describe cell lines producing six variant forms of deminectin corresponding to variant forms of fibronectin produced by alternative splicing. In constructing fibronectin cDNAs encoding the six variant forms, we also made use of the ability of retroviral vectors to generate cDNAs by accurate splicing of cloned genomic segments. These constructs should be valuable in analyses of the structure-function relationships of fibronectins.