Objective To identify the gene encoding the long-chain fatty acid transport protein (FATP) of Dermatophagoides farinae and its molecular characteristics. Methods According to the transcriptome sequencing data of D. farinae, we obtained the FATP-coding sequence for designing primers. Then RT-PCR was used to amplify the full-length gene fragment from total RNA of D. farinae, followed by construction of the prokaryotic expression plasmid pET28a(+)-FATP. After obtaining FATP sequence successfully, the structure and function of amino acid sequence were analyzed by using the software of Expasy、singaIP5.0、GOR4 and TMpred, and performed homology analysis through Clustal Omega and MEGA-X. Results The full length of the gene coding sequence was 1 071 bp. The encoded protein was composed of 356 amino acids. It was soluble, with a grand average of hydropathicity of -0.39, and had a stable structure. The secondary structure included alpha-helix (25.56%), extended strand (23.04%), and random coil (51.40%). The protein had two transmembrane regions, The transmembrane area from the outside to the inside is 1-20 aa, and the transmembrane area from the inside to the outside is 100-116 aa. The evolutionary tree derived from homologous amino acid sequences showed that the sequence of D. farinae was clustered with that of silkworm. Conclusion The full length of the FATP-coding gene and the molecular characteristics of FATP are identified in D. farinae, laying a foundation for exploring its physiological phenomena and developing control measures.
ZHU Han-ting, WU Mei-li, ZHOU Ying, WU Bing, CUI Yu-bao
. Cloning and sequencing of the long-chain fatty acid transporter gene of Dermatophagoides farinae[J]. Chinese Journal of Vector Biology and Control, 2020
, 31(2)
: 169
-174
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DOI: 10.11853/j.issn.1003.8280.2020.02.010
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