Objective To establish a DNA barcoding technique for the molecular identification of flies based on COⅠ gene and to make up for the deficiency of morphological identification of flies. Methods Sixteen flies commonly found at Yantai Port were used as subjects in this study. After morphological identification, total DNA was extracted from the single hind foot of each fly. According to reference, the international COⅠ primer LCO1490: 5′-GGT CAA CAA ATC ATA AAG ATA TTG G-3′ and HCO2198: 5′-TAA ACT TCA GGG TGA CCA AAA AAT CA-3′ were used to set up a PCR system. Sequence of COⅠ was successfully amplified, followed by sequencing and aligning of purified PCR product. Phylogenetic study of COⅠ gene was performed using Mega 5.05 by neighbor-joining (NJ), maximum parsimony (MP), and maximum likelihood (ML) methods. Results The amplified COⅠ fragments of sixteen flies were 690 bp in size, with an average A＋T content of 68.53%. The sequence alignment and phylogenetic study showed that the results of COⅠ gene-based identification were consistent with morphological identification. Conclusion The DNA barcoding technique established in this study is able to amplify COⅠ gene from flies and thereby identify flies with the same result as morphological identification; this technique is particularly suitable for the identification of a trace amount of samples.
WANG Ying, LU Min, LI Jin-qing, LIU Ming-jie, YUE Qiao-yun
. Study on rapid molecular identification of vector flies based on COⅠ gene[J]. Chinese Journal of Vector Biology and Control, 2014
, 25(5)
: 427
-431
.
DOI: 10.11853/j.issn.1003.4692.2014.05.011
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