Jie Yu-Kun, Luo Zhi-Ping, Xie Jia-Wei, Cheng Chang-Hong, Ma Hong-Ling, Liu Guang-Xin, Jiang Jian-Jun, Deng Yi-Qin, Feng Juan, Guo Zhi-Xun
Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, 510300, PR China; National Demonstration Center for Experimental Fisheries Science Education, Shanghai Engineering Research Center of Aquaculture, Shanghai Ocean University, Shanghai, 201306, China.
Zhuhai Modern Agriculture Development Center, Zhuhai, Guangdong, 519070, PR China.
Fish Shellfish Immunol. 2022 May;124:39-46. doi: 10.1016/j.fsi.2022.03.042. Epub 2022 Mar 31.
Phosphofructokinase (PFK), the key enzyme of glycolysis, can catalyze the irreversible transphosphorylation of fructose-6-phosphate forming fructose-1, 6-biphosphate. In the present study, a PFK gene from the mud crab Scylla paramamosain, named SpPFK, was cloned and characterized. The full length of SpPFK contained a 5'untranslated region (UTR) of 249 bp, an open reading frame of 2,859 bp, and a 3'UTR of 1,248 bp. The mRNA of SpPFK was highly expressed in the gill, followed by the hemocytes and muscle. The expression of SpPFK was significantly up-regulated after mud crab dicistrovirus-1 (MCDV-1) infection. Knocking down SpPFK in vivo by RNA interference significantly reduced the expression of lactate dehydrogenase after MCDV-1 infection. Furthermore, silencing of SpPFK in vivo increased the survival rate of mud crabs and decreased the MCDV-1 copies in the gill and hepatopancreas after MCDV-1 infection. All these results suggested that SpPFK could play an important role in the process of MCDV-1 proliferation in mud crab.
磷酸果糖激酶（PFK）是糖酵解的关键酶，可催化6-磷酸果糖不可逆地转磷酸化形成1,6-二磷酸果糖。在本研究中，克隆并鉴定了来自拟穴青蟹的一个PFK基因，命名为SpPFK。SpPFK的全长包含一个249 bp的5'非翻译区（UTR）、一个2859 bp的开放阅读框和一个1248 bp的3'UTR。SpPFK的mRNA在鳃中高表达，其次是血细胞和肌肉。拟穴青蟹双顺反子病毒-1（MCDV-1）感染后，SpPFK的表达显著上调。通过RNA干扰在体内敲低SpPFK可显著降低MCDV-1感染后乳酸脱氢酶的表达。此外，在体内沉默SpPFK可提高拟穴青蟹的存活率，并降低MCDV-1感染后鳃和肝胰腺中MCDV-1的拷贝数。所有这些结果表明，SpPFK可能在MCDV-病毒在拟穴青蟹中的增殖过程中发挥重要作用。
