Wei Quande, Yu Xinbing, Lim Ho Sung, Cha Hyung Joon
Department of Pathogenic Biology, Medical College of Sun Yat-sen University, Guangzhou 510089, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2006 Jun;23(3):609-14.
In order to determine whether H1 histone proteins are associated with innate immune antimicrobial response in goldfish, we extracted the total RNA from the hemocytes of goldfish (Carassius auratus), designed 3 pairs of primers based on the previous antimicrobial peptide sequences from fish and performed RT-PCR. Among 3 obtained-PCR products, we identified a novel histone H1 coding sequence of 576 bp which belongs to the histone H1 family and is 78% homologous with the amino acid sequence of histone H1 from Salmon salar that had been found with an important role in salmon defenses against infectious pathogens. The H1 histone of goldfish contained 3 predicting cleavage sites that divided the protein into 4 parts. We successfully cloned and expressed the whole CDs (No ATG) of H1 histone and its N-terminal part (2-38aa) in Pichia pPIC9K expression system. The products of H1 histone and its N-terminal deriving peptide (AEVAPAASAPPAKAPKKKSAAKAKKAGPAVGDLIVKA) show antimicrobial activity. The results suggested that the H1 histone fragment reported in this paper is a novel antimicrobial peptide found in goldfish. H1 histone plays an important role in innate immune responses of goldfish.
为了确定H1组蛋白是否与金鱼的先天性免疫抗菌反应相关，我们从金鱼（Carassius auratus）的血细胞中提取了总RNA，根据先前鱼类抗菌肽序列设计了3对引物，并进行了RT-PCR。在获得的3个PCR产物中，我们鉴定出一个576 bp的新型组蛋白H1编码序列，它属于组蛋白H1家族，与大西洋鲑组蛋白H1的氨基酸序列同源性为78%，而大西洋鲑组蛋白H1在鲑鱼抵御传染性病原体方面发挥着重要作用。金鱼的H1组蛋白包含3个预测的切割位点，将该蛋白分为4部分。我们在毕赤酵母pPIC9K表达系统中成功克隆并表达了H1组蛋白的完整编码区（无ATG）及其N端部分（2-38aa）。H1组蛋白及其N端衍生肽（AEVAPAASAPPAKAPKKKSAAKAKKAGPAVGDLIVKA）的产物具有抗菌活性。结果表明，本文报道的H1组蛋白片段是在金鱼中发现的一种新型抗菌肽。H1组蛋白在金鱼的先天性免疫反应中发挥着重要作用。
