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Expression and alternative splicing analysis of a large-conductance calcium-activated potassium channel gene in Plutella xylostella.
Archives of Insect Biochemistry and Physiology ( IF 1.5 ) Pub Date : 2020-06-17 , DOI: 10.1002/arch.21720 Jie Xu 1 , Hai-Hao Ma 2 , Zhe-Ming Liu 2 , Wei Zheng 2 , Xiao-Yi Lai 2 , Hang Zhu 2 , Jia Liu 2 , Yong Zhou 2 , Xiao-Mao Zhou 1, 2
Archives of Insect Biochemistry and Physiology ( IF 1.5 ) Pub Date : 2020-06-17 , DOI: 10.1002/arch.21720 Jie Xu 1 , Hai-Hao Ma 2 , Zhe-Ming Liu 2 , Wei Zheng 2 , Xiao-Yi Lai 2 , Hang Zhu 2 , Jia Liu 2 , Yong Zhou 2 , Xiao-Mao Zhou 1, 2
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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.
中文翻译：
小菜蛾大电导钙激活钾通道基因的表达和可变剪接分析。
大电导钙激活钾通道（BK Ca）在调节昆虫神经回路和运动中起着重要作用，因此是杀虫剂的潜在目标。在这项研究中，发现BK Ca的抑制剂iberiotoxin延长了小菜蛾幼虫上乙酸乙酯的麻醉时间。因此，克隆了编码BK Ca的α亚基的慢戳基因的编码序列，以研究该通道在小菜蛾中的功能，并表征了其在发育阶段和组织中的基因表达谱。pxslo的总长度DNA超过19.9 kb，具有四个备选剪接位点（ASP-A，ASP-C，ASP-E和ASP-G），并且pxslo的编码序列具有最高的剪接频率（GenBank ID：MN938456）是3,405个碱基对。表征的PxSlo蛋白包含先前在其他昆虫中鉴定的保守结构域。定量逆转录聚合酶链反应分析表明，pxslo在小菜蛾的所有发育阶段都有表达，在成年人中最高。在幼虫阶段，pxslo主要在头部和表皮中表达，而有限的蛋白质在中肠中表达。在成人阶段，pxslo在头中高表达，其次在卵巢小管中表达，而在睾丸或翅膀中不表达。这些结果表明，BK Ca在小菜蛾中起着重要的生理作用，并为功能研究和BK Ca抑制剂的筛选提供了有用的信息。
更新日期：2020-06-17
中文翻译：
小菜蛾大电导钙激活钾通道基因的表达和可变剪接分析。
大电导钙激活钾通道（BK Ca）在调节昆虫神经回路和运动中起着重要作用，因此是杀虫剂的潜在目标。在这项研究中，发现BK Ca的抑制剂iberiotoxin延长了小菜蛾幼虫上乙酸乙酯的麻醉时间。因此，克隆了编码BK Ca的α亚基的慢戳基因的编码序列，以研究该通道在小菜蛾中的功能，并表征了其在发育阶段和组织中的基因表达谱。pxslo的总长度DNA超过19.9 kb，具有四个备选剪接位点（ASP-A，ASP-C，ASP-E和ASP-G），并且pxslo的编码序列具有最高的剪接频率（GenBank ID：MN938456）是3,405个碱基对。表征的PxSlo蛋白包含先前在其他昆虫中鉴定的保守结构域。定量逆转录聚合酶链反应分析表明，pxslo在小菜蛾的所有发育阶段都有表达，在成年人中最高。在幼虫阶段，pxslo主要在头部和表皮中表达，而有限的蛋白质在中肠中表达。在成人阶段，pxslo在头中高表达，其次在卵巢小管中表达，而在睾丸或翅膀中不表达。这些结果表明，BK Ca在小菜蛾中起着重要的生理作用，并为功能研究和BK Ca抑制剂的筛选提供了有用的信息。
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