title: Proteoglycans: gene cloning.
abstract: Aggrecan is a large proteoglycan that plays roles in numerous tissues during vertebrate development and adult life. The 6,327-nt chick aggrecan coding sequence had been determined from overlapping clones, but a full-length cDNA, needed for use in transgenic expression studies, had not been constructed. The strategy employed to do so was to generate two overlapping cDNA subfragments that shared a unique restriction site in the overlap and then join them at that site. These subfragments were obtained and cloned into the TOPO-TA vector pCR2.1. Digestion of the two constructs with the shared-site enzyme, XbaI, produced vector/5'-cDNA and 3'-cDNA fragments with XbaI-ends; these were ligated to produce the final full-length cDNA.
authors.affiliation: Departments of Pediatric, The University of Chicago, Chicago, IL, USA.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Cortes
authors.forename: James R
authors.initials: JR
authors.lastname: Mensch
authors.forename: Miriam
authors.initials: M
authors.lastname: Domowicz
authors.forename: Nancy B
authors.initials: NB
authors.lastname: Schwartz
mesh_terms: D053674:Aggrecans / Q000235:genetics*; D000818:Animals; D002645:Chickens; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D051379:Mice
chemical_list: D053674:Aggrecans; D018076:DNA, Complementary