Objective To clone the ultraviolet sensitive opsin (UV-opsin) gene of Aedes albopictus, to characterize the sequences of UV-opsin gene and its encoded protein, and to provide a molecular basis for the study of its biological function. Methods The degenerate primers were designed according to the conserved sequences of the UV-opsin gene from mosquitoes, and the UV-opsin gene of Ae. albopictus was amplified by RT-PCR. The PCR product was purified and ligated into the pET28a(+) vectors with EcoRⅠ digestion and XhoⅠ to construct a prokaryotic expression plasmid. The positive recombinant plasmid was identified by PCR and sequenced. Structural features and function were analyzed and predicted using Expasy, Protparam, Tmpred, and ScanProsite. Sequence homology was analyzed by Clustal X 1.81 and a phylogenetic tree was constructed using MEGA 6.0 software. Results The cDNA sequence of Ae. albopictus UV-opsin was obtained, which was 1 146 bp in full length (including termination codon) and encoded 381 amino acid residues. The predicted molecular weight was 42 500 and the isoelectric point was 7.06. The homology was 99.74% with Ae. albopictus (XP_029730343) and 96.85% with Ae. aegypti (ABF18478). The phylogenetic tree showed that insects from the same genera and orders clustered together. Aedes was clustered with the genus Culex, and then with Anopheles. This UV-opsin had 7 transmembrane topology domains, 1 G-protein-coupled receptor, 4 N-myristoylation sites, 3 N-glycosylation sites, 1 tyrosine kinase phosphorylation site, 3 protein kinase C phosphorylation sites, and 2 casein kinase Ⅱ phosphorylation sites. The UV-opsin contained conserved motifs LRTPSN, DRY and QAKK, which were critical for its function. Conclusion The UV-opsin gene sequence of Ae. albopictus is cloned, which has the structural characteristics of opsins. The results provide a foundation for further research on the function of Ae. albopictus UV-opsin gene.
GUO Xiu-xia, CHENG Peng, LIU Li-juan, WANG Hai-fang, ZHANG Chong-xing, WANG Huai-wei, GONG Mao-qing
. Cloning and sequence features of ultraviolet sensitive opsin gene in Aedes albopictus[J]. Chinese Journal of Vector Biology and Control, 2021
, 32(5)
: 553
-559
.
DOI: 10.11853/j.issn.1003.8280.2021.05.009
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