MOLECULAR-CLONING AND EXPRESSION OF MOUSE PROCALCITONIN

The nucleotide sequence for mouse calcitonin was determined from a cDN A obtained using a polymerase chain reaction (PCR) based method, the r apid amplification of cDNA ends (RACE)-PCR. Primers designed from high ly conserved regions in the coding sequences of known rat and human ca lcitonin cDNAs were used to amplify calcitonin cDNA as 5'-end and 3'-e nd fragments from mouse thyroid RNA. The obtained cDNA is 850 bp in le ngth most probably representing the entire mouse calcitonin mRNA, It c ontains an open reading frame coding for a 136 amino acid protein with a calculated M(r) of 15,143. Comparison of the deduced amino acid seq uences of preprocalcitonin of mice with other species revealed highest homologies to the rat (93%) and human (77%) sequences. A recombinant form of mouse procalcitonin (rmPCT) of approximately 17 kDa was expres sed as a fusion peptide in E.coli transformed with a PCR-cloned expres sion construct. (C) 1996 Academic Press, Inc.