| 大肠杆菌免疫刺激DNA的克隆及其对抗体产生的影响 |
| |
| 引用本文： | 李爱芹,庄文忠,任红梅,钱凤芹.大肠杆菌免疫刺激DNA的克隆及其对抗体产生的影响[J].中国预防兽医学报,2002,24(3):178-181. |
| |
| 作者姓名： | 李爱芹 庄文忠 任红梅 钱凤芹 |
| |
| 作者单位： | 山东省农业科学院生命科学研究中心,山东,济南,250100 |
| |
| 摘 要： | 利用聚合酶链式反应（Polymerase Chain Reaction,PCR）从大肠杆菌基因组中扩增免疫刺激基因，并把扩增的基因片段克隆和pGEM－T载体。PstI酶切结果表明重组质粒（rP）均含有扩增的基因片段；rP1与rP2是相同的重组质粒。利用酶联免疫吸附检测了重组质粒对小鼠产生抗体的影响，结果显示：重组质粒1与牛血清白蛋白（BSA），免疫小鼠能产生较同的抗体水平，而重组质粒3则对小鼠产生的BSA抗体水平没有影响。对重组质粒1的序列分析,克隆的基因片段中富含CpG序列，并含有数个具有强烈免疫刺激作用的RRCGYY（如：AACGTT）序列。本研究为基因免疫机理的深入研究和基因免疫佐剂的开发奠定了基础。
|
| 关 键 词： | 大肠杆菌 免疫刺激 对抗体 DNA克隆 细菌基因组 菌种 酶 |
| 文章编号： | 1008-0589(2002)03-0178-04 |
| 修稿时间： | 2001年6月28日 |
Molecular Cloning and Immune Effects of Bacterial Immunostimulatory DNA |
| |
| LI Ai_qin,ZHUANG Wen_zhong ,REN Hong_mei,QIAN Feng_qin.Molecular Cloning and Immune Effects of Bacterial Immunostimulatory DNA[J].Chinese Journal of Preventive Veterinary Medicine,2002,24(3):178-181. |
| |
| Authors: | LI Ai_qin ZHUANG Wen_zhong REN Hong_mei QIAN Feng_qin |
| |
| Institution: | LI Ai_qin,ZHUANG Wen_zhong *,REN Hong_mei,QIAN Feng_qin |
| |
| Abstract: | Bacterial DNA contains a much higher frequency of unmethylated CpG dinucleotides,which have strong immunostimulating effects in inducing activation of B cells,NK cells and monocytes.Previous study suggested that CpG DNA may be an excellent immune adjuvant candidate.For cloning bacterial immunostimulatory DNA,in this research,polymerase chain reaction(PCR)was used to amplify immunostimulatory sequence from bacterial genomic DNA.Amplified three DNA fragments were cloned into pGEM_T plasmid.Results of digesting recombinant plasmid(rP)with PstI demonstrated that the rP1 and rP2 was the same one.Effect of the recombinant plasmid(rP)to antibody production was tested by immunizing mice with mixture of the rP and bovine serum albumin(BSA) or BSA alone as a control.The antibody titer was detected by enzyme_linked immunosorbent assay,the results of which showed that rP1 could enhance level of antibody production,but the rP3 could not.Sequence analysis of the gene fragment in rP1 demonstrated that the DNA contained abundant CG dinucleotides and several RRCGYY sequences. |
| |
| Keywords: | PCR CpG Plasmid Antibody |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
