Kim Ju-Won, Choi Hye-Sung, Kwon Mun-Gyeong, Park Myoung-Ae, Hwang Jee-Youn, Kim Do-Hyung, Park Chan-Il
Department of Marine Biology & Aquaculture, Institute of Marine Industry, College of Marine Science, Gyeongsang National University, 455, Tongyeong 650-160, Republic of Korea.
Pathology Division, National Fisheries Research and Development Institute, Busan 619-900, Republic of Korea.
Results Immunol. 2011 Aug 26;1(1):45-52. doi: 10.1016/j.rinim.2011.08.002. eCollection 2011.
Natural killer cell enhancing factor (NKEF) belongs to the defined peroxiredoxin (Prx) family. Rock bream NKEF cDNA was identified by expressed sequence tag (EST) analysis of rock bream liver that was stimulated with the LPS. The full-length RbNKEF cDNA (1062 bp) contained an open reading frame (ORF) of 594 bp encoding 198 amino acids. RbNKEF was significantly expressed in the gill, liver, and intestine. mRNA expression of NKEF in the head kidney was examined under viral and bacterial challenge via real-time RT-PCR. Experimental challenge of rock bream with Edwardsiella tarda, Streptococcus iniae, and RSIV resulted in significant increases in RbNKEF mRNA in the head kidney. To obtain a recombinant NKEF, the RbNKEF ORF was expressed in Escherichia coli BL21 (DE3), and the purified soluble protein exhibited a single band corresponding to the predicted molecular mass. When kidney leucocytes were treated with a high concentration of rRbNKEF (10 μg/mL), they exhibited significantly enhanced cell proliferation and viability under oxidative stress.
自然杀伤细胞增强因子（NKEF）属于已明确的过氧化物还原酶（Prx）家族。通过对经脂多糖刺激的真鲷肝脏进行表达序列标签（EST）分析，鉴定出了真鲷NKEF cDNA。真鲷NKEF cDNA全长1062 bp，包含一个594 bp的开放阅读框（ORF），编码198个氨基酸。真鲷NKEF在鳃、肝脏和肠道中显著表达。通过实时逆转录聚合酶链反应（RT-PCR）检测了在病毒和细菌攻击下，头肾中NKEF的mRNA表达。用迟缓爱德华氏菌、海豚链球菌和真鲷虹彩病毒对真鲷进行实验性攻击，导致头肾中真鲷NKEF mRNA显著增加。为了获得重组NKEF，将真鲷NKEF开放阅读框在大肠杆菌BL21（DE3）中表达，纯化后的可溶性蛋白呈现出一条与预测分子量相对应的单条带。当用高浓度的重组真鲷NKEF（10 μg/mL）处理肾白细胞时，它们在氧化应激下表现出显著增强的细胞增殖和活力。
