| 摘要: |
| 为了解山羊内在防御系统中发挥的重要作用,在山羊防御素保守序列的基础上设计了12条不同理化性质的防御素(DGBD1~12)基因,克隆到毕赤酵母表达载体pPICZαA的EcoR I和Not I酶切位点,构建重组表达载体pPICZαA-DGBD。经SaI线性化后电击转化毕赤酵母GS115,经PCR鉴定阳性重组菌。经过摇瓶发酵和甲醇诱导,阳性重组菌的发酵上清液对大肠杆菌,金黄色葡萄球菌等具有较强的抑制作用。最小抑菌浓度测定显示重组质粒对大肠杆菌的抑制作用最强。研究结果显示,疏水指数较高的序列DGBD1~DGBD3的抗菌活性较好,稳定指数较低的序列DGBD4~DGBD6抗菌活性仅次于疏水指数,说明DGBD结构稳定性和疏水性对其抗菌活性有较大影响。 |
| 关键词: β-防御素 内在防御系统 理化性质 基因工程 抗菌活性 |
| DOI：10.11841/j.issn.1007-4333.2015.01.016 |
| 投稿时间：2014-04-10 |
| 基金项目:国家肉羊产业技术体系建设项目(CARS39-1A) |
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| Effects of protein physicochemical properties of goat β-defensins on their antimicrobial activity |
|
KANG Jian-feng1, XU Ping-ping1, LIU Gui-qiong1, YANG Jian-xiong2, JIANG Xun-ping1
|
| (1.College of Animal Sciences and Technology, Huazhong Agricultural University, Wuhan 430070, China;2.School of Veterinary Medicine, Northeast Agricultural University, Harbin 150030, China) |
| Abstract: |
| Goat β-defensin-1 (GBD1) is a class of cationic peptides widely present in goat intestines,liver,lungs and reproductive tract and other tissues.GBD1 plays an important role in the goat intrinsic defense system.In this study,goat defensins were designed based on conserved sequences for 12 different physical and chemical properties of defensins (DGBD1-12) and cloned Pichia expression vectors pPICZA in EcoR I and Not I,and recombinant expression vectors.After linearized by SaI,it was electroporated into Pichia pastoris GS115,and identified by PCR of bacteria.After fermentation and methanol induction,recombinant bacteria fermentation supernatant Escherichia coli,Staphylococcus aureus had a strong inhibitory effect.Determination of minimum inhibitory concentration of E.coli recombinant plasmid displayed the strongest inhibition.The results of this study show that DGBD structural stability and hydrophobicity have a greater impact on their antibacterial activity. |
| Key words: β-defensins intrinsic defense system physical and chemical properties genetic engineering antibacterial activity |
