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源于草鱼肠道抗菌肽的重组表达、分离纯化和抗菌活性分析
发表时间：2012-06-15 浏览量：1650 下载量：423
| 全部作者： | 史玉婷，孙科军，陶虚谷，苏建明，曹蓉，陈韬 |
| 作者单位： | 湖南农业大学动物医学院；湖南农业大学动物科技学院 |
| 摘 要： | 根据草鱼肠道中分离得到的抗菌肽（antibacterial peptide, AP）氨基酸测序结果，人工合成草鱼抗菌肽基因，并将该合成基因克隆至pET32a载体中，构建成含有6×His标签的重组质粒pET- AP. 重组质粒转化至大肠杆菌BL21中，经异丙基硫代半乳糖苷（IPTG）诱导后，进行十二烷基硫酸钠- 聚丙烯酰胺凝胶电泳（SDS- PAGE）分析。测序结果表明：插入片段的序列和位置均正确。通过Ni2+- NTA柱亲和层析纯化，用肠激酶将纯化的重组蛋白中的His标签切除后回收重组抗菌肽，采用滤纸片方法测定其抗菌活性。实验结果表明，重组抗菌肽能明显抑制大肠杆菌的生长。 |
| 关 键 词： | 兽医学；抗菌肽；重组表达；纯化；抗菌活性 |
| Title： | Expression, purification and antimicrobial activity analysis of antimicrobial peptide originated from intestine of grass carp |
| Author： | SHI Yuting, SUN Kejun, TAO Xugu, SU Jianming, CAO Rong, CHEN Tao |
| Organization： | Veterinary Faculty, Hunan Agricultural University; College of Animal Science and Technology, Hunan Agricultural University |
| Abstract： | According to the amino acid sequence of intestine of grass carp, antibacterial peptide (AP) coding genes were synthesized artificially and cloned into vector pET32a to construct a recombinant plasmid termed pET- AP which contains a 6譎is tag. Plasmid pET- AP was transferred to E.coli strain BL21. SDS- PAGE analysis of recombinant protein was carried out after induced by IPTG. Sequencing results indicated that the sequence and location of insert fragment in pET- AP were correct. Recombinant protein was purified by Ni2+- NTA column affinity chromatography. And enterokinase was used to recover the recombinant AP. Antimicrobial activity assay showed that recombinant AP could inhibit the growth of E.coli. |
| Key words： | veterinary medicine; antibacterial peptide; recombinant expression; purification; antimicrobial activity |
| 发表期数： | 2012年6月第11期 |
| 引用格式： | 史玉婷，孙科军，陶虚谷，等. 源于草鱼肠道抗菌肽的重组表达、分离纯化和抗菌活性分析[J]. 中国科技论文在线精品论文，2012，5（11）：1081-1086. |
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