Cloning of a cDNA complementary to rat preputial gland [beta]-glucuronidase mRNA

Messenger RNA was isolated from rat preputial glands by guanidine HCl extraction, ethanol and salt precipitation, followed by chromatography on oligo(dT) cellulose. Double-stranded cDNA was synthesized from the mRNA and inserted into the Pst 1 site of the plasmid pBR322 by the poly(dG)[middle dot]poly(dC) tailing and annealing procedure. The hybrid plasmids were used to transform HB101. Recombinant clones were screened for those containing cDNA inserts complementary to [beta]-glucuronidase mRNA by a hybridization-selection procedure. One clone, containing an insert of about 1.2 kilobases, hybridized to preputial gland mRNA which, when translated , gave a product that migrated with the [beta]-glucuronidase subunit on polyacrylamide gels.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/24054/3/0000305.pd