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| 嗜水气单胞菌溶血素A表达、免疫原性及活性检测 | |
| 引用本文： | 赵秀敏,高勤学,陈鸿军,韩先干,丁铲.嗜水气单胞菌溶血素A表达、免疫原性及活性检测[J].中国兽医寄生虫病,2011,19(1):45-50. |
| 作者姓名： | 赵秀敏 高勤学 陈鸿军 韩先干 丁铲 |
| 作者单位： | 1. 河南农业职业学院,郑州,451450 2. 江苏畜牧兽医职业技术学院,泰州,225300 3. 中国农业科学院上海兽医研究所,上海,200241 |
| 基金项目： | 中央公益性科研院所基础研究基金项目 |
| 摘 要： | 根据已发表的嗜水气单胞菌（Aeromonas hydrophila,Ah）溶血素A基因序列设计一对特异性引物,以TPS30菌株基因组为模板,通过PCR扩增溶血素A基因,经T-A克隆、序列测定和分析,结果表明,该基因包含1482 bp碱基,编码494个氨基酸。将溶血素A定向克隆至表达载体pET32a（＋）中,构建重组表达质粒pET-THA,在IPTG诱导下成功获得重组表达蛋白His-GroEL,大小为68 kDa。对该质粒在大肠杆菌BL21（DE3）中的表达特性分析表明,最佳的诱导表达条件为：在0.1 mmol/L的IPTG浓度下,21℃诱导表达3 h。表达的溶血素A蛋白纯化后免疫小鼠,经Western blot检测,表明该蛋白具有免疫原性。对纯化的溶血素A蛋白复性后,进行溶血实验,结果表明该蛋白具有溶血活性。通过本实验的研究,为进一步研究嗜水气单胞菌溶血素的生物学功能奠定了基础。 |
| 关 键 词： | 嗜水气单胞菌 溶血素 克隆 表达 大肠杆菌 |
EXPRESSON, IMMUNOGENICITY AND HEMOLYTIC ACTIVITY OF AEROMONAS HYDROPHILA HEMLYSIN A |
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| ZHAO Xiu-min,GAO Qin-xue,CHEN Hong-jun,HAN Xian-gan,DING Chan.EXPRESSON, IMMUNOGENICITY AND HEMOLYTIC ACTIVITY OF AEROMONAS HYDROPHILA HEMLYSIN A[J].Chinese Journal of Veterinary Parasitology,2011,19(1):45-50. | |
| Authors: | ZHAO Xiu-min GAO Qin-xue CHEN Hong-jun HAN Xian-gan DING Chan |
| Institution: | ZHAO Xiu-min1,GAO Qin-xue2,CHEN Hong-jun3,HAN Xian-gan3,DING Chan3 (1.Henan Vocational College of Agriculture,Zhengzhou 451045,China,2.Jiangsu Animal Husbandry Veterinary College,Taizhou 225300,3.Shanghai Veterinary Research Institute CAAS,Shanghai 200241,China) |
| Abstract: | A pair of specifical primers was designed according to gene sequence of Aermonas hydrophila hemolysin A which has been reported.Using Aermonas hydrophila strain TPS30 genomic DNA as template,Aermonas hydrophila hemolysin A was amplied by PCR and cloned into pGEM-T vector.The result of sequence analysis shown that the length of hemolysin A is 1482 bp,encoding a 494-amino-acid protein.This gene was cloned into pET-32a(+) vector and generated recombinant express plasmid pET-THA,a recombinant protein His-GroEL ... |
| Keywords: | Aeromonas hydrophila hemolysin cloning expression E coli |
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