RESUMEN
Objective To construct cDNA library from adults of Anopheles dirus for cloning the immune genes or related genes for malaria parasites development. Methods The mRNA of adult Anopheles dirus was isolated. The library was constructed by using the Zap Express vector(Stratagene) and the quality was evaluated. Results The efficiency of the library was 2.1?10 6 pfu/ml with 98% clones positive. The average length of the insert fragment was over 1 kb. Conclusion cDNA library of adult Anopheles dirus with high efficiency can be constructed by using the Zap Express library construction Kit (Stratagene).
RESUMEN
Objective To clone the partial cDNA sequence of prophenoloxidase (PPO) gene of Anopheles dirus . Methods Degenerative primers were designed according to the conserved sequence blocks within the prophenoloxidase of insects. RNA sequence of the larva of Anopheles dirus was amplified by RT PCR to get the prophenoloxidase cDNA which was then cloned into T vector and sequenced. The partial cDNA sequence of prophenoloxidase gene was analysed and compared with other prophenoloxidase gene of insects. Results The partial cDNA sequence of AdPPO4 was 597 bp, and its deduced amino acid sequence was 199aa. The cDNA sequence homology and amino acid sequence homology was 84% and 90%, respectively, as compared with the PPO4 gene of Anopheles gambiae . Conclusion The AdPPO4, with high sequence homology with the PPO4 gene of Anopheles gambiae , is successfully cloned from the larva of Anopheles dirus .