| 家蚕抗菌肽moricin在大肠杆菌中的融合表达及抗菌活性测定 |
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| 引用本文： | 赵岩龙,沈兴家,郭锡杰.家蚕抗菌肽moricin在大肠杆菌中的融合表达及抗菌活性测定[J].蚕业科学,2008,34(2):232-236. |
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| 作者姓名： | 赵岩龙 沈兴家 郭锡杰 |
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| 作者单位： | 江苏科技大学,江苏,镇江,212003;江苏科技大学,江苏,镇江,212003;中国农业科学院蚕业研究所,农业部家蚕生物技术重点开放实验室,江苏,镇江,212018 |
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| 基金项目： | 国家重点基础研究发展计划(973计划)
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国家高技术研究发展计划(863计划)
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江苏省高技术研究计划(农业)项目 |
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| 摘 要： | Moricin是家蚕中发现的一种抗菌肽,对革兰氏阴性和阳性菌都有较强的抗菌活性,具有良好的开发应用前景。为了建立一种大量表达和快速分离moricin的方法,采用PCR拼接法获得moricin基因后重组到表达载体pET-32M上,并在大肠杆菌(E.coli)中融合表达。表达产物存在于上清中为可溶状态,经Ni-NTA柱纯化获得融合蛋白,再经凝血酶(thrombin)酶切后过2次Ni-NTA柱获得纯度为90%以上的moricin,液相测定法表明纯化获得的moricin对大肠杆菌具有抗菌活性。
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| 关 键 词： | 家蚕 抗菌肽 Moricin 融合表达 抗菌活性 |
Fusion Expression and Test of Antibacterial Activity of the Antibacterial Peptide Moricin of Bombyx mori in Escherichia coil |
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| ZHAO Yan-Long,SHEN Xing-Jia,GUO Xi-Jie.Fusion Expression and Test of Antibacterial Activity of the Antibacterial Peptide Moricin of Bombyx mori in Escherichia coil[J].Acta Sericologica Sinica,2008,34(2):232-236. |
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| Authors: | ZHAO Yan-Long SHEN Xing-Jia GUO Xi-Jie |
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| Abstract: | Moricin is a novel antibacterial peptide from the silkworm (Bombyx mori) with activities against fungi and gram-positive and gram-negative bacteria. In order to establish an over-expressed and facilitated purification method, the moricin gene was obtained by a recursive PCR strategy using five overlap oligonucleotides and cloned into the expression vector pET-32M and expressed in Escherichia.coli BL21. The expressed product was soluble in supernatant and the fusion protein was purified by Ni-NTA. After digested with thrombin and purified with other 2 times of Ni-NTA, moricin with a purity of over 90 percent was obtained. Liquid turbidity analysis indicated that the purified moricin exhibited the antibacterial activity. |
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| Keywords: | Bombyx mori Antibacterial peptide Moricin Fusion expression Antibacterial activity |
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