Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki, Gakuen Kibanadai-nishi, Miyazaki, Japan.
Lett Appl Microbiol. 2010 Jan;50(1):112-9. doi: 10.1111/j.1472-765X.2009.02763.x.
In the present study, we have cloned a new family of anti-lipopolysaccharide factor (ALF) from haemocytes of kuruma shrimp Marsupenaeus japonicus (MjALF2) using RACE method.
Transcriptional analysis of MjALF2 gene in the organs of healthy shrimp revealed prominent expression in gills and muscle. In vitro LPS stimulation in the lymphoid organ cells resulted in significant increase in expression at 48, 8 and 12 h poststimulation, compared to the nonstimulated cells. In vivo injection of V. penaeicida does not show any high expression in time course assay. Phylogenetic analysis showed MjALF2 is placed in the group closer to P. monodon isoform 1 and 2 than to MjALF1. The full-length MjALF2 gene consists of 558 bp with a 363 -bp open reading frame, encoding 121 amino acids. The deduced peptide contains a putative signal peptide of 22 amino acids with molecular mass of about 13.8 kDa molecular mass. The deduced amino acid sequence of MjALF2 showed 83.3 and 56.7% identity with ALF sequences of P. monodon.
The present work revealed the presence of MjALF2 gene, which is highly expressed in gills and muscle of healthy kuruma shrimp. Further studies are required to clarify the involvement of MjALF2 in immune responses for using as a therapeutic agent.
Antimicrobial peptides are promising potential therapeutic agents for disease control in aquaculture. Understanding the relation of MjALF2 with innate immunity mechanism will lead to develop better health management strategies for long-term sustainability of the shrimp industry.
本研究采用 RACE 方法，从日本对虾（Marsupenaeus japonicus）血细胞中克隆了一个新的抗脂多糖因子（ALF）家族（MjALF2）。
健康虾各组织器官中 MjALF2 基因的转录分析显示，其在鳃和肌肉中表达最为显著。在淋巴器官细胞体外 LPS 刺激下，与未刺激细胞相比，刺激后 48、8 和 12 小时表达显著增加。在体内注射 V. penaeicida 的时间过程检测中，没有显示出任何高表达。系统发育分析表明，MjALF2 与 P. monodon 同工型 1 和 2 比与 MjALF1 更接近。全长 MjALF2 基因由 558bp 组成，其中包含一个 363bp 的开放阅读框，编码 121 个氨基酸。推导的肽含有一个推定的 22 个氨基酸的信号肽，分子量约为 13.8kDa。MjALF2 的推导氨基酸序列与 P. monodon 的 ALF 序列具有 83.3%和 56.7%的同一性。
本研究揭示了 MjALF2 基因的存在，该基因在健康日本对虾的鳃和肌肉中高度表达。需要进一步的研究来阐明 MjALF2 在免疫反应中的作用，以便将其作为一种治疗剂使用。
抗菌肽是水产养殖疾病控制有前途的潜在治疗剂。了解 MjALF2 与先天免疫机制的关系将为虾类产业的长期可持续发展制定更好的健康管理策略。
