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Characterization and in vitro expression of arginine kinase gene in the invasive western flower thrips, Frankliniella occidentalis

Author:
Dong, Fan, Zhang, Nan, Xie, Zhijuan, Meng, Xiangkun, Qian, Kun, Ji, Caihong, Lu, Mingxing, Du, Yuzhou, Wang, Jianjun
Source:
Comparative biochemistry and physiology 2019 v.229 pp. 51-57
ISSN:
1096-4959
Subject:
Escherichia coli, Frankliniella occidentalis, arginine, arginine kinase, complementary DNA, energy metabolism, enzyme activity, enzyme kinetics, exons, gene expression, genomics, insecticides, insects, introns, messenger RNA, nucleotides, open reading frames, prepupae, quantitative polymerase chain reaction, reverse transcriptase polymerase chain reaction, screening, sequence analysis
Abstract:
Arginine kinase (AK) plays a critical role in insect energy metabolism and has been proposed to be a potential insecticide target for commercial exploitation. In this study, the full length cDNA encoding a typical group 1 insect AK (FoAK) was isolated from the western flower thrips (WFT), Frankliniella occidentalis (Pergande). Sequence analysis showed that FoAK contains an open reading frame of 1068 nucleotides, which encods a protein of 355 amino acid residues including the signature sequence pattern of ATP-guanidino kinases. Genomic structure analysis showed that the coding region of FoAK contains five exons connected by four introns. RT-qPCR analysis revealed that the mRNA expression of FoAK was developmentally regulated with the lowest level in prepupal stage. Enzymatic activity analysis of the recombinant enzymes expressed in Escherichia coli showed that FoAK is highly stereo specific for L-arginine versus D-arginine and the apparent Michaelis constant for L-arginine (KmArg) is comparable to that of AKs from a variety of species. This research should enable further investigation of the function as well as in vitro screening for inhibitors of FoAK.
Agid:
6293523