State Key Laboratory of Marine Environmental Science, College of Ocean & Earth Sciences, Xiamen University, Xiamen, Fujian 361102, PR China.
Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources, Xiamen University, Xiamen, Fujian 361102, PR China; State-Province Joint Engineering Laboratory of Marine Bioproducts and Technology, Xiamen University, Xiamen, Fujian 361102, PR China.
Comp Biochem Physiol A Mol Integr Physiol. 2019 Jan;227:25-31. doi: 10.1016/j.cbpa.2018.08.017. Epub 2018 Sep 7.
The early developmental stages of the mud crab Scylla paramamosain suffer from high mortality caused by pathogen infections; however, few immune associated factors are known. Lipopolysaccharide and β-1,3-glucan-binding protein (LGBP) functions as a typical pathogen recognition receptor and plays an important role in the innate immune system of invertebrates. In this study we characterized a LGBP gene (SpLGBP) which was highly expressed in the late embryonic, zoea I larval stage and hepatopancreas of S. paramamosain.. It encodes 364 amino acids, composed of several conserved domains like the bacterial glucanase motif. The recombinant SpLGBP protein (rSpLGBP) was obtained through the E.coli expression system, in which two 6◊His-tags were added to both C and N terminals during vector construction for the improvement of purification efficiency. In vivo the study showed that the SpLGBP mRNA was significantly up-regulated under Vibrio parahaemolyticus and a lipopolysaccharide (LPS) challenge in the hemocytes and hepatopancreas. The ELISA binding assay in vitro indicated that the rSpLGBP was capable of binding to LPSs and peptidoglycan (PGN). The rSpLGBP could agglutinate both G+ and G- bacteria in the presence of Ca. Our results suggest that SpLGBP may play an immunological role against pathogenic infection in the early developmental stages of S. paramamosain.
锯缘青蟹（Scylla paramamosain）早期胚胎发育阶段易受病原体感染而导致高死亡率，但人们对其免疫相关因子知之甚少。脂多糖和β-1,3-葡聚糖结合蛋白（LGBP）作为一种典型的病原体识别受体，在无脊椎动物的先天免疫系统中发挥着重要作用。本研究中，我们对锯缘青蟹 LGBP 基因（SpLGBP）进行了特征描述，该基因在晚期胚胎、Zoea I 幼虫期和肝胰腺中高表达。它编码 364 个氨基酸，由几个保守结构域组成，如细菌葡聚糖酶基序。通过大肠杆菌表达系统获得了重组 SpLGBP 蛋白（rSpLGBP），在载体构建过程中，在 C 和 N 末端添加了两个 6◊His 标签，以提高纯化效率。体内研究表明，SpLGBP mRNA 在副溶血弧菌和脂多糖（LPS）刺激的血细胞和肝胰腺中显著上调。体外 ELISA 结合试验表明，rSpLGBP 能够与 LPS 和肽聚糖（PGN）结合。rSpLGBP 能够在 Ca 存在下凝集 G+和 G-细菌。我们的研究结果表明，SpLGBP 可能在锯缘青蟹早期发育阶段对致病性感染发挥免疫作用。
