We will clone and express the cystic fibrosis transmembrane conductance regulator (CFTR) in insect and mammalian cells. We will develop methods to purify these two CFTRs, which will differ only in their extent of glycosylation, and establish omgeneity by 1D- and 2D-gel electrophoresis, amino acid composition, and N-terminal amino acid sequencing. the purified function of the CFTRs, proper glycsoylation and phosphorylation, development of methods for large-scale purification for use in diagnostic assays and replacement therapy.
