Construction of Molecular Phylogeny for Mosquitoes using the Sequences of Mitochondrial ND5 and ITS-2 in Ribosomal DNA

• Main Authors: Chang-Ta Yang, 楊正大
• Other Authors: Chau-Mei Ho
• Format: Others
• Language: zh-TW
• Published: 2003
• Online Access: http://ndltd.ncl.edu.tw/handle/98164458963194678896

碩士 === 國立陽明大學 === 寄生蟲學研究所 === 91 === Sequences of the internal transcribed spacer 2 (ITS-2) of ribosomal DNA and partial mitochondrial ND5 were used to construct the phylogenetic dendrograms 27 species of mosquitoes in the genera of Aedes, Anopheles, Armigerus, Culex, Triptonoides, and Toxorhynchites collected in Taiwan in this study. DNA fragments of either ITS-2 or partial ND5 were amplified by polymerase chain reaction (PCR) with specific primers, and were cloned into vector of PUC18 T. The sequences of the inserts were obtained from the plasmid DNA containing appropriate inserted DNA fragment. The length of the ITS-2 of 27 species ranges from 249 bp to 533 bp, with the GC content of 44.8 to 57.2%. However, the DNA fragment of DN5 was derived from 15 species of mosquitoes. The length of the DN5 fragments in these mosquitoes ranged from 553 bp to 576 bp with GC content of 17- 20.6%. All obtained nucleotide sequences of ITS-2 and ND5 were aligned by Pileup in SeqWeb. Pairwise distances were measured using the program of Molecular Evolution Genetic Analysis (MEGA). Neighbor-joining (NJ) trees and maximum parasimony trees were derived from the distance matrix with MEGA. Results of our study showed that the sequences of ND 5 appeared to be more reasonable than that of ITS-2 at the level above species. However, the sequence of ITS-2 is useful in the aspect of identify subspecies of Culex pipiens and the forms of Anopheles minimums.