剑尾鱼IgM基因的克隆及免疫对其组织表达的影响  

Cloning and Expression of IgM in Tissues of Immunized Xiphophorus helleri

作　　者：韩进刚[1] 付小哲[1] 石存斌[1] 李凯彬[1] 潘厚军[1] 吴淑勤[1] 

机构地区：[1]中国水产科学研究院珠江水产研究所

出　　处：《广东海洋大学学报》2007年第6期1-6,共6页Journal of Guangdong Ocean University

基　　金：科技部社会公益项目(2004DIB1J029);国家科技支撑计划项目(2006BAD03B05)

摘　　要：以免疫两周的剑尾鱼为材料,提取脾脏总RNA,通过逆转录-聚合酶链式反应(RT-PCR)扩增出分泌型免疫球蛋白M(secretory immunoglobulin M,sIgM)重链基因的核心序列,再应用3’和5’快速扩增cDNA末端(RACE)方法扩增其末端序列,拼接后获得剑尾鱼sIgM重链全长cDNA序列。全长序列含有免疫球蛋白的信号序列:FKCIANH,得到的5个可变区序列可能来源于4个不同的VH家族。经用半定量RT-PCR分析sIgM在脾脏和头肾的表达变化,初步认为利用剑尾鱼头肾sIgM的mRNA表达变化替代检测血清抗体效价具有可能性。The total RNA was extracted from spleen of Xiphophorus helleri which had been immunized for two weeks. The core sequence was amplified by RT-PCR. The cDNA end sequences were amplified by RACE, and then the full- length cDNA of sIgM heavy chains gene were obtained by putting all of them. The full-length sequence contains the immunoglobulin signature sequence： FKCIANH. The five sequences, encoding sIgM variable region, were likely to root in the four VH families. The expressions of sIgM, in spleen and head kidney ofXiphophorus helleri, were analyzed by semi-quantitative RT-PCR. According to the results, it was possible to use the semi-quantitative RT-PCR method to detect the change of slgM expression in head kidney instead of the potency of sIgM antibody in serum, and provide the theory foundation for the immune model of aquatic laboratory animal.

关 键 词：剑尾鱼 免疫球蛋白M 基因表达:基因克隆 

分 类 号：Q78[生物学—分子生物学] Q959.475