Jiang Jingwei, Zhou Zunchun, Dong Ying, Chen Zhong, Sun Hongjuan, Yang Aifu, Gao Shan, Su Hesheng
Liaoning Key Lab of Marine Fishery Molecular Biology, Liaoning Ocean and Fisheries Science Research Institute, Dalian, Liaoning 116023, PR China.
Liaoning Key Lab of Marine Fishery Molecular Biology, Liaoning Ocean and Fisheries Science Research Institute, Dalian, Liaoning 116023, PR China.
Fish Shellfish Immunol. 2017 Aug;67:11-18. doi: 10.1016/j.fsi.2017.05.061. Epub 2017 May 26.
In order to preliminarily illustrate the functional differences of phenoloxidases (POs) in Apostichopus japonicus, the full-length cDNAs of two POs (named as AjPOⅡ and AjPOⅢ, respectively) were cloned from the coelomocytes of A. japonicus using 3'- and 5'-rapid amplification of cDNA ends method, and combined with the previously acquired full-length cDNA of a laccase-type PO from A. japonicus (Accession No. KF040052, named as AjPOⅠ), the sequence structure and phylogenic status of POs from A. japonicus (AjPOs) were comparatively analyzed, and the transcriptional expression of AjPOs in different tissues, at different developmental stages and after different bacterial challenges was determined with quantitative real-time PCR method. Sequence analysis indicated AjPOⅡ and AjPOⅢ were both laccase-type POs, coincident to the results of phylogenic analysis. Sequence analysis also showed that AjPOⅠ had a transmembrane domain (J. Jiang et al., 2014), AjPOⅡ contained a signal peptide, and AjPOⅢ possessed a signal peptide and a transmembrane domain, implying that three AjPOs might play different roles in immune and physiological processes. Transcriptional expression analysis showed that AjPOⅡ and AjPOⅢ were most abundant in tube feet, while AjPOⅠhad the highest expression level in coelomocytes (J. Jiang et al., 2014), suggesting that AjPOⅠ may be mainly involved in immune response, while AjPOⅡ and AjPOⅢ are probably responsible for other physiological processes in addition to immune response. Besides, three AjPOs were determined to have different expression patterns during organism development and different spectrums of response against bacteria, which further indicated that there might be immune and physiological functional differentiation among three AjPOs.
为初步阐明刺参中酚氧化酶（POs）的功能差异，采用cDNA末端快速扩增法从刺参体腔细胞中克隆了两种POs的全长cDNA（分别命名为AjPOⅡ和AjPOⅢ），并结合先前获得的刺参漆酶型PO的全长cDNA（登录号KF040052，命名为AjPOⅠ），对刺参POs（AjPOs）的序列结构和系统发育地位进行了比较分析，并用实时定量PCR法测定了AjPOs在不同组织、不同发育阶段及不同细菌刺激后的转录表达。序列分析表明，AjPOⅡ和AjPOⅢ均为漆酶型POs，与系统发育分析结果一致。序列分析还显示，AjPOⅠ有一个跨膜结构域（Jiang等人，2014年），AjPOⅡ含有一个信号肽，AjPOⅢ具有一个信号肽和一个跨膜结构域，这意味着三种AjPOs可能在免疫和生理过程中发挥不同作用。转录表达分析表明，AjPOⅡ和AjPOⅢ在管足中含量最高，而AjPOⅠ在体腔细胞中的表达水平最高（Jiang等人，2014年），这表明AjPOⅠ可能主要参与免疫反应，而AjPOⅡ和AjPOⅢ除免疫反应外可能还负责其他生理过程。此外，确定三种AjPOs在生物发育过程中具有不同的表达模式，对细菌的反应谱也不同，这进一步表明三种AjPOs之间可能存在免疫和生理功能分化。
