Characterization and in vitro expression of arginine kinase gene in the invasive western flower thrips, Frankliniella occidentalis
Characterization and in vitro expression of arginine kinase gene in the invasive western flower thrips, Frankliniella occidentalis
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入侵性西花蓟马（Frankliniella occidentalis）精氨酸激酶基因的特征及体外表达
DOI：
10.1016/j.cbpb.2019.01.003
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发表时间：
2019-03-01
影响因子：
2.2
通讯作者：
Wang, Jianjun
中科院分区：
文献类型：
--
作者：
Dong, Fan;Zhang, Nan;Wang, Jianjun
精氨酸激酶（AK）在昆虫的能量代谢中起着重要作用，已被认为是一种潜在的商业开发的杀虫剂靶标。本研究从西花蓟马Frankliniella oceanentalis（Pergande）中克隆了一个编码第1组昆虫AK（FoAK）基因的全长cDNA。序列分析表明，FoAK含有一个1068个核苷酸的开放阅读框，编码355个氨基酸，具有ATP-胍基激酶的特征序列。基因组结构分析表明，FoAK基因编码区由5个外显子和4个内含子组成。RT-qPCR分析表明，FoAK基因的表达受发育调控，在蛹前期表达量最低。在大肠杆菌中表达的重组酶的酶活性分析表明，FoAK是高度立体特异性的L-精氨酸与D-精氨酸和L-精氨酸的表观米氏常数（Km（Arg））是可比的从各种物种的AK。这项研究应使进一步调查的功能，以及在体外筛选抑制剂的FoAK。
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 (Km(Arg)) 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.