金鱼H 1组蛋白基因及其N端片段的克隆、表达及抗菌活性  

Cloning,Expression and Antibacterial Activity of Histone H1 and Its N-terminal Peptide from Carassius auratus

作　　者：魏泉德[1] 余新炳[1] Lim Ho Sung Cha Hyung Joon 

机构地区：[1]中山大学医学院病原生物学室,广州510089 [2]韩国蒲项工科大学化学工程系暨分子与生命科学部

出　　处：《生物医学工程学杂志》2006年第3期609-614,共6页Journal of Biomedical Engineering

摘　　要：用RT-PCR方法从金鱼体内成功分离得到编码H 1组蛋白的基因,基因登录号AY 184811。该基因与来源于大西洋鲑鱼的抗菌蛋白-H 1组蛋白有极高的同源性(78%)。将全长基因及其N端(2-38氨基酸)序列成功克隆于毕赤酵母分泌表达载体pP IC 9K并表达;表达产物初步检测有抗菌活性。结果表明,金鱼H 1组蛋白为一种新发现的抗菌蛋白,其N端来源肽(AEVAPAA SAPPAKAPKKK SAAKA KKAGPAVGDL IVKA)为抗微生物肽。金鱼H 1组蛋白及其来源肽在金鱼先天免疫防御中发挥作用。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.

关 键 词：抗微生物肽 H1组蛋白 金鱼 毕赤酵母 RT—PCR 

分 类 号：R346[医药卫生—基础医学]