Molecular cloning, characterization and tissue expression of prophenoloxidase cDNA from the mosquito Armigeres subalbatus inoculated with Dirofilaria immitis microfilariae

• Published in: Insect molecular biology 1998-02, Vol.7 (1), p.31-40
• Main Authors: Cho, W. L., Liu, H. S., Lee, C. H., Kuo, C. C., Chang, T. Y., Liu, C. T., Chen, C. C.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Blotting, Northern; Catechol Oxidase - genetics; Cloning, Molecular; Culicidae - enzymology; Culicidae - genetics; Culicidae - parasitology; Dirofilaria immitis - physiology; DNA, Complementary; Dogs; Enzyme Precursors - genetics; Female; In Situ Hybridization; Molecular Sequence Data; Sequence Analysis, DNA; Sequence Homology, Amino Acid
• ISSN: 0962-1075; 1365-2583
• DOI: 10.1046/j.1365-2583.1998.71049.x

A cDNA encoding mosquito Armigeres subalbatus pro‐phenol oxidase (As‐pro‐PO) was obtained by rapid amplification of cDNA ends‐polymerase chain reaction (RACE‐PCR) after Dirofilaria immitis inoculation. The 2205 bp As‐pro‐PO cDNA contains a 32 bp 5′‐noncoding region, a 2055 bp open reading frame (685 amino acids), and a 118 bp 3′‐noncoding region. Hydrophobic signal peptide for the endoplasmic reticulum targeting is not found in the NH2‐terminal region. Two potential copper‐binding domains, amino acids 197–245 and 345–412, are highly homologous to those of the other insect pro‐POs. A 2.2 kb As‐pro‐PO transcript was identified by Northern blot analysis using D. immitis microfilariae‐inoculated A. subalbatus. Both in situ hybridization and Northern blot analysis demonstrated that As‐pro‐PO mRNA was synthesized in mosquito haemocytes but not in other tissues, i.e. fat bodies, midguts and ovaries, etc.