DETECTION OF MESSENGER-RNA ENCODING AN ANTIBACTERIAL-METALLOPROTEIN (MPII) BY IN-SITU HYBRIDIZATION WITH A CDNA PROBE GENERATED BY POLYMERASE CHAIN-REACTION IN THE WORM NEREIS-DIVERSICOLOR

Based on partial amino acid sequence of an antibacterial-metalloprotei n (MPII) consisting of a N-terminal fragment(1-33 th) and an 43 amino acids long internal fragment, two oligonucleotides primers were synthe sized and used to generate a cDNA fragment by the PCR method. The spec ificity of the PCR synthesized 220 bp fragment was verified by hybrid- arrest translation and sequencing. In situ hybridization performed wit h this cDNA fragment on Nereis diversicolor whole body defined two spe cific sites of recognition: a cluster of cells floating in the coelom and two types of muscles (perineural and oblique). Finally, the chrono logical expression of the MPII was postulated.