论文部分内容阅读
从草鱼Ctenopharyngodon idella肝肾cDNA文库中克隆得到胶原凝集素基因。草鱼胶原凝集素全长cDNA为1128bp,其中5′非编码区229bp,3′非翻译区104bp,最大开放阅读框为795bp,编码264个氨基酸。系统进化分析表明草鱼胶原凝集素与斑马鱼的亲缘关系最近。根据草鱼胶原凝集素序列特征,克隆了包含糖基识别域(CRD)的cDNA,并进行原核表达、纯化获得其重组蛋白PCRD。进行PCRD与6种细菌的凝集和糖抑制实验,结果表明半乳糖、葡萄糖、甘露糖和麦芽糖4种糖都会使PCRD与嗜水气单胞菌的凝集明显下降甚至极大地干扰凝集;麦芽糖使金黄色葡萄球菌的凝集明显下降,而肽聚糖和甘露糖会使凝集受到抑制;此外,PCRD的凝集反应不依赖Ca2+。
The collagen lectin gene was cloned from the grass carp Ctenopharyngodon idella liver-kidney cDNA library. Grass carp collagen lectin full-length cDNA was 1128bp, including 5 ’non-coding region 229bp, 3’ untranslated region 104bp, the largest open reading frame is 795bp, encoding 264 amino acids. Phylogenetic analysis showed that grass carp collagen lectin was the most closely related to zebrafish. According to the characteristics of the collagen lectin sequence of grass carp, the cDNA containing the glycosylation domain (CRD) was cloned and prokaryotic expressed. The recombinant protein PCRD was obtained. PCRD and 6 kinds of bacteria agglutination and sugar inhibition experiments, the results show that four kinds of sugar galactose, glucose, mannose and maltose PCRD and Aeromonas hydrophila coagulation decreased significantly or even greatly interfere with agglutination; maltose gold Agglutination of Staphylococcus aureus decreased significantly, while peptidoglycan and mannose inhibited the agglutination. In addition, the agglutination reaction of PCRD was independent of Ca2 +.