Expression and alternative splicing analysis of a large-conductance calcium-activated potassium channel gene in Plutella xylostella.
Expression and alternative splicing analysis of a large-conductance calcium-activated potassium channel gene in Plutella xylostella.
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DOI：
10.1002/arch.21720
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发表时间：
2020-06
影响因子：
2.2
通讯作者：
Jie Xu;Haihao Ma;Zheming Liu;W. Zheng;Xiaoyi Lai;Hang Zhu;Jia Liu;Yong Zhou;Xiaomao Zhou
中科院分区：
文献类型：
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作者：
Jie Xu;Haihao Ma;Zheming Liu;W. Zheng;Xiaoyi Lai;Hang Zhu;Jia Liu;Yong Zhou;Xiaomao Zhou
大电导钙激活钾通道（BKCa）在昆虫神经回路和运动的调节中发挥着重要作用，因此是杀虫剂的潜在靶标。本研究发现，BKCa抑制剂伊比利亚毒素可延长乙酸乙酯对小菜蛾幼虫的麻醉时间。因此，克隆了编码BKCaα亚基的slowpoke基因的编码序列，以研究该通道在小菜蛾中的功能，并对其在发育阶段和组织中的基因表达谱进行了表征。 pxslo DNA总长度超过19.9 kb，包含4个可变剪接位点（ASP-A、ASP-C、ASP-E和ASP-G），剪接频率最高的pxslo编码序列（GenBank ID：MN938456）为3,405个碱基对。表征的 PxSlo 蛋白包含先前在其他昆虫中发现的保守结构域。定量逆转录聚合酶链反应分析表明，pxslo在小菜蛾的各个发育阶段都有表达，其中在成虫中表达量最高。在幼虫阶段，pxslo主要在头部和表皮表达，而中肠表达有限的蛋白。在成年阶段，pxslo在头部高度表达，其次是卵巢小管，而在睾丸和翅膀中不表达。这些结果表明BKCa在小菜蛾中发挥着重要的生理作用，为BKCa抑制剂的功能研究和筛选提供了有用的信息。
The large-conductance calcium-activated potassium channel (BKCa ) plays an important role in the regulation of insect neural circuits and locomotion, and thus is a potential target of insecticides. In this study, iberiotoxin, an inhibitor of BKCa , was found to prolong the anesthetic time of ethyl acetate on Plutella xylostella larvae. Therefore, the coding sequence of slowpoke gene coding the alpha subunit of BKCa was cloned to investigate the function of this channel in P. xylostella, and the gene expression profile in the developmental stages and tissues was also characterized. The total length of pxslo DNA was more than 19.9 kb, which harbored four alternative splicing sites (ASP-A, ASP-C, ASP-E, and ASP-G), and the coding sequence of pxslo with the highest frequency of splicing (GenBank ID: MN938456) was 3,405 base pair. The characterized PxSlo protein contained conserved domains previously identified in other insects. Quantitative reverse transcription-polymerase chain reaction analysis showed that pxslo was expressed in all the developmental stages of P. xylostella, with the highest level in adults. In the larval stage, pxslo was mainly expressed in the head and epidermis, while a limited protein was expressed in the midgut. In the adult stage, pxslo was highly expressed in the head, followed by in the ovarian tubule, and was not expressed in the testis or wings. These results suggest that BKCa plays an important physiological role in P. xylostella and provides useful information for the functional study and screening of BKCa inhibitors.