Detection of mRNA 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-metalloprotein (MPII) consisting of a N-terminal fragment (1-33th) and an 43 amino acids long internal fragment, two oligonucleotides primers were synthesized and used to generate a cDNA fragment by the PCR method. The specificity of the PCR synthesized 220 bp fragment was verified by hybrid-arrest translation and sequencing. In situ hybridization performed with this cDNA fragment on Nereis diversicolor whole body defined two specific sites of recognition: a cluster of cells floating in the coelom and two types of muscles (perineural and oblique). Finally, the chronological expression of the MPII was postulated.
Salzet-Raveillon
B
B
Laboratoire de Phylogénie Moléculaire des Annélides ERS CNRS 20, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France.
Rentier-Delrue
F
F
Dhainaut
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D001483:Base Sequence; D003001:Cloning, Molecular; D015342:DNA Probes; D017403:In Situ Hybridization; D008667:Metalloproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D011077:Polychaeta / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics*
D000900:Anti-Bacterial Agents; D015342:DNA Probes; D008667:Metalloproteins; D012333:RNA, Messenger