| 猪源抗菌肽PBD-1在毕赤酵母中的表达及鉴定 |
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| 引用本文： | 江学斌,陈嘉蔚,杨军,刘顺枝,肖安吉,卢志鹏,楚品品,黄朝远,蔡海明,马苗鹏,张玲华.猪源抗菌肽PBD-1在毕赤酵母中的表达及鉴定[J].中国畜牧兽医,2016,43(3):644-649. |
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| 作者姓名： | 江学斌 陈嘉蔚 杨军 刘顺枝 肖安吉 卢志鹏 楚品品 黄朝远 蔡海明 马苗鹏 张玲华 |
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| 作者单位： | 1. 广州大学生物工程研究所, 广州 510006;2. 华南农业大学生命科学学院, 广东省农业生物蛋白质功能与调控重点实验室, 广州 510642 |
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| 基金项目： | 广州市属高校科技计划项目(1201420755) |
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| 摘 要： | 本试验旨在实现猪β防御素1 (poricine-β-defensin 1,PBD-1)在毕赤酵母中的表达,获得有抗菌活性的抗菌肽PBD-1。根据PBD-1的氨基酸序列和酵母密码子偏好性,设计优化其核苷酸序列,利用SOE-PCR技术获得PBD-1基因序列,克隆到毕赤酵母表达载体pPIC9K中,构建重组质粒pPIC9K-PBD-1,经SacⅠ线性化后转入毕赤酵母SMD1168中。PCR筛选得到阳性酵母表达菌株,经甲醇诱导后得到分子质量约4.5 ku的抗菌肽PBD-1。抗菌特性研究结果表明,表达产物抗菌肽PBD-1对大肠杆菌、金黄色葡萄球菌及枯草芽孢杆菌均有较好的抑制效果。
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| 关 键 词： | PBD-1基因 表达 毕赤酵母 PCR |
| 收稿时间： | 2015-08-17 |
Expression and Identification of Porcine Antimicrobial Peptide PBD-1 in Pichia pastoris |
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| JIANG Xue-bin,CHEN Jia-wei,YANG Jun,LIU Shun-zhi,XIAO An-ji,LU Zhi-peng,CHU Pin-pin,HUANG Chao-yuan,CAI Hai-ming,MA Miao-peng,ZHANG Ling-hua.Expression and Identification of Porcine Antimicrobial Peptide PBD-1 in Pichia pastoris[J].China Animal Husbandry & Veterinary Medicine,2016,43(3):644-649. |
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| Authors: | JIANG Xue-bin CHEN Jia-wei YANG Jun LIU Shun-zhi XIAO An-ji LU Zhi-peng CHU Pin-pin HUANG Chao-yuan CAI Hai-ming MA Miao-peng ZHANG Ling-hua |
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| Institution: | 1. Research Institute of Bioengineering of Guangzhou University, Guangzhou 510006, China;2. Function and Regulation of Agricultural Biotechnology Protein of Guangdong Province Key Laboratory, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China |
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| Abstract: | In order to produce antimicrobial peptide PBD-1 with bioactivity in Pichia pastoris,according to published amino acid sequence of PBD-1 and the partiality codon of yeast,the PBD-1 gene was amplified by SOE-PCR and cloned into pPIC9K to construct a recombinant expression vector pPIC9K-PBD-1.The recombinant vector was linearized by SacⅠ,and then transformed into SMD1168 by electroporation.Positive yeast expression strain was obtained by PCR.The antimicrobial peptide PBD-1 (approximately 4.5 ku) was expressed by methanol induction.Antibacterial activity assay showed that the antimicrobial peptide PBD-1 had better antibacterial activity against E.coli,S.aureus and B.subtilis. |
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| Keywords: | PBD-1 gene expression Pichia pastoris PCR |
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