CLONING EXPRESSION AND PURIFICATION OF CFTR

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. $ = TOTAL AWARD AMTS & NOT LIMITED TO PORTION OF PROJECT RELATED TO SUBJECT OF SEARCH SUBPROJECT $ = TOTAL AWARD AMOUNT DIVIDED BY NUMBER OF SUBPROJECTS SOURCE: CRISP FORMAT F FY 97 LAST UPDATE 04-07-98 1QUERY 1536 ID SEARCH 06/01/98 PAGE 274 --PROJECT NUMBER......1 R43 DK53142-01 INVESTIGATOR NAME/ADDRESS FY 97 KISLAUSKIS, EDWARD H IRG/INTRAMURAL UNIT..ZRG2 BIOMEDICAL RESEARCH MODELS INC AWARD AMOUNT......... $85,254 12 NANCY DRIVE RUTLAND, MA 01543-1717 PERFORMING ORGANIZATION: BIOMEDICAL RESEARCH MODELS, INC. TITLE CRYOPRESERVATION OF THE BB/WOR RAT ABSTRACT: The overall goal in Phase I is to develop a method to efficiently cryopreserve embryos from the BB/Wor rat strains. Funding is requested to determine which established technique (slow equilibration, two step equilibration, or vitrification) is most suited to obtain cryopreserved embryos. Once a method has been selected, we will cryopreserve circa 20 years of genetic inbreeding and concurrently make available new reagents for the study of human diabetic embryopathy.