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The terminal structure plays an important role in the biological activity of cecropin CMIV

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Science China(Life Sciences) 1999年第5期42卷 494-500页

作者：窦非谢维董雪吟徐贤秀 State Key Laboratory of Pharmaceutical Biotechnology Department of Biochemistry Nanjing University Nanjing China

Antibacterial peptides have received increasing attention as a new pharmaceutical substance. But the molecular mechanism of lysis is still poorly understood. CMIV gene and mutant CMIV gene in GST fusion system were expressed. After cleaving with different cleavage reagents, the peptide with an excess of N-terminus and with an un-amidated C-terminus stopped the activity while the peptide with an excess Asn at the C-terminus had the activity level the same as natural CMIV. The results showed that the terminal structure of cecropin CMIV played an important role in its biological activity.

关键词： cecropin mutant GST antibacterial activity terminal structure.

Fusion expression of mutated cecropin CMIV in E. coli

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Science China(Life Sciences) 1997年第3期40卷 225-231页

作者：谢维邱奇峰董雪吟华子春徐贤秀 State Key Laboratory of Pharmaceutical Biotechnology Department of Biochemistry Nanjing University Nanjing 210093 China

A cDNA coding mutated cecropin CMIV from Bombyx mori was synthesized according to its amino acid sequense using E .coli biased codons .The gene was cloned into the fusion expression vector pEZZ318 and was expressed in E .coli *** fusion protein produced was purified by affinity chromatography to yield 26 mg/L fusion product .The anti-bacterial activities of recombinant cecropin CMIV were recovered after cleavage by chemical method.

关键词： cecropin fusion expression E. coli.

无载体主干序列的bar和cecropin B基因表达框共转化水稻

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Acta Genetica Sinica 2003年第2期30卷 135-141页

作者：赵艳于彦春钱前颜美仙黄大年浙江大学生命科学学院中国水稻研究所生物工程系农业部水稻生物学重点实验室杭州310006 中国水稻研究所生物工程系农业部水稻生物学重点实验室

基因枪等直接转化法可将去除质粒载体主干序列的外源基因表达框导入植物基因组 ,消除质粒主干序列对植物基因组的影响 ,同时由于转基因分子较小 ,比较容易实现多个基因的共转化 ,在植物基因工程育种上有重要的应用价值。研究了基因枪介... 详细信息

基因枪等直接转化法可将去除质粒载体主干序列的外源基因表达框导入植物基因组 ,消除质粒主干序列对植物基因组的影响 ,同时由于转基因分子较小 ,比较容易实现多个基因的共转化 ,在植物基因工程育种上有重要的应用价值。研究了基因枪介导外源基因表达框 (包括启动子、基因开放阅读框和终止子 )转化水稻的影响因素和转基因的整合模式 ,结果表明 :(1)基因枪介导外源基因表达框转化水稻的频率约在 0 1%～ 0 5 %之间 ,非选择标记基因与选择标记基因的共转化频率约为 5 0 %～ 6 0 % ,增加基因表达框DNA浓度可提高转化率。相同基因构建物对不同品种水稻的转化率不同 ,基因构建物的侧边序列对基因枪转化的品种差异可能具有重要影响。 (2 )非选择标记基因cecropinB表达框在水稻基因组内整合模式简单 ,仅有 1～ 3个拷贝 ;筛选标记基因bar表达框却比完整质粒转化后的整合模式复杂得多 ,插入拷贝数在 4～ 14个之间。线形基因表达框的游离DNA末端和CaMV

关键词：无载体主干序列 bar cecropin B基因表达框共转化水稻

The Cytotoxic Effect of cecropin A and cecropin B on the MDA-MB-231 and M14K Tumour Cell Lines

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Journal of Biomedical Science and Engineering 2014年第8期7卷 504-515页

作者： Radu Anghel Daniela Jitaru Laurentiu Badescu Manuela Ciocoiu Magda Badescu Department of Cell and Molecular Biology University of Medicine and Pharmacy “Grigore T. Popa” Iasi Romania Department of Pathophysiology University of Medicine and Pharmacy “Grigore T. Popa” Iasi Romania Laboratory ofMolecular Biology Regional Institute of Oncology Iasi Romania

The aim of the present in vitro study was to assess the tumoricidal potential of the following natural peptides belonging to the cecropin family, namely cecropin A and B, on a series of tumour cell lines: MDA-MB-231 (breast adenocarcinoma) and M14K (human mesothelioma). The experimental results reveal that the cytotoxic effects of the two peptides depend on their concentration. Their efficiency is significant at 120 μM concentrations and it persists even at 60 μM concentrations. The effects were insignificant at 30 μM concentrations. On the other hand, the cytotoxic potential was not significantly dependant on the type of peptide but more on the type of tumour cell line used. The MDA MB 231 line cells were much more sensitive to the action of cecropins A and B than the M14K line cells. The prospects brought about by this experimental research consist of the collection of in vitro experimental data on the tumoricidal potential of these natural cytotoxic peptides on tumour cells. This will enable specialists to develop future in vivo experimental models in order to test the antitumor effect of these cytotoxic peptides. The ultimate goal would be the discovery of agents with efficient antitumor properties, i.e. with maximum tumoricidal effects and minimum toxic side effects.

关键词： Breast Adenocarcinoma cecropin A cecropin B Human Mesothelioma Tumoricidal Potential

Introduction of cecropin B gene into rice (Oryza sativa L.) by particle bombardment and analysis of transgenic plants

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Science China(Life Sciences) 1996年第6期39卷 652-661页

作者：黄大年朱冰杨炜薛锐肖晗田文忠李良材戴顺洪 China National Rice Research Institute Hangzhou 310006 China Institute of Genetics Chinese Academy of Sciences Beijing 100101 China

An expression vector pCBl suitable for rice transformation, harboring a synthesized cecropin B gene and a selectable marker gene (bar), was constructed. It was introduced into immature embryos of two japonica varieties by particle bombardment, and several transgenic plants were obtained. The results from Basta treatment, PCR analysis, dot and Southern blot analysis of cecropin B gene in transgenic plants indicated that both bar and cecropin B gene were integrated into the genome of transformed plants. Northern blot analysis of transgenic plants showed the expression of cecropin B gene at transcriptional level. Some of transgenic plants revealed improved resistances to two types of bacterial diseases, rice bacterial blight and rice bacterial streak to different extent.

关键词： cecropin B gene particle bombardment bar gene rice (Oryza sativa L..).

Study of high-level expression of Musca domestica cecropin with partner molecular in Escherichia coli

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Study of high-level expression of Musca domestica cecropin w...

致倦库蚊cecropin基因家族的比较和系统发育分析

格莱姆抗菌肽——抗菌肽开发与应用技术研讨会

作者： Wang wei,Zheng Xueli*(Department of Pathogenic Biology,School of Public Health and Tropical Medicine,Southern Medical University,Guangzhou 510515,China)

Objective To utilize partner molecular for effective expressing Musca domestica cecropin,research that partner molecular influences expression of *** cecropin in ***. Methods Gene of mature cecropin and ubiquitin of *** were amplified by *** bioinformatics methods,we analyzed secondary structure of the fusion protein of thioredoxin-MC and thioredoxin-ubiquitin-MC’s *** expression plasmids of pET32a-MC and pET32a-Ubiquitin -MC were *** expression plasmids were transformed into *** and were induced with IPTG *** of protein expression of pET32a-MC and pET32a-Ubiquitin-MC by SDS-PAGE was finished Results The protein quantity of expression of pET32a-Ubiquitin&MC were significantly higher than that of *** Profile of mature peptide of cecropin was transformed by *** fusion protein of thioredoxin-MC isn’t toxic for host *** results showed the partner molecule ubiquitin can significantly result in high-level expression of peptide antibiotic of M. demostica cecropin.

关键词： Musca domestica cecropin Partner molecular Fusion expression Ubiquitin

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基因组学与应用生物学 2016年第2期35卷 294-301页

作者：赵文静张晶张春林翟素珍王吉平贵州医科大学基础医学院生物学教研室贵阳550004

利用生物信息学方法比较分析了致倦库蚊cecropin家族4成员的基因结构、氨基酸序列、蛋白理化特性,预测蛋白空间结构,并基于7种昆虫绘制分子进化树,随后使用RT-PCR方法获取致倦库蚊cecropin家族4成员的c DNA。结果表明：致倦库蚊Cecropi... 详细信息

利用生物信息学方法比较分析了致倦库蚊cecropin家族4成员的基因结构、氨基酸序列、蛋白理化特性,预测蛋白空间结构,并基于7种昆虫绘制分子进化树,随后使用RT-PCR方法获取致倦库蚊cecropin家族4成员的c DNA。结果表明：致倦库蚊cecropin家族基因有两条正向编码,两条反向编码,长度为330~544 bp,由2个外显子和1个内含子构成;氨基酸序列长度为59~61 aa,具有两条基序和多个保守位点;编码的蛋白理论分子量为6 284.6~6 619.0 k D,p I值为10.36~11.24,亲水性平均系数为0.393~0.551;蛋白预测的二级结构和三级结构结果基本一致,它们具有1个信号肽结构域和1个催化结构域,结构域主要由α-螺旋构成;7种昆虫的cecropin蛋白进化关系与物种进化关系基本一致,并以“目”或“亚目”为单位分别聚类,CPIJ005108与致倦库蚊cecropin祖先基因最相似。本研究的分析结果可为蚊科cecropin结构和功能的研究提供有价值的信息。

关键词：致倦库蚊 cecropin 基因家族生物信息学

抗菌肽cecropin及其在转基因植物抗菌中的应用

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中国生物工程杂志 2008年第5期28卷 122-127页

作者：李波张耀洲浙江理工大学生命科学学院生物化学研究所杭州310018

cecropin抗菌肽,又名天蚕素,是一类具有抗菌能力的小分子多肽,热稳定性好,抗菌谱广,在各个应用领域已经得到广泛的研究。就cecropin抗菌肽结构与功能的关系、作用机制和其在转基因植物抗菌领域中的应用进行综述,阐述了转基因植物表达Cec... 详细信息

cecropin抗菌肽,又名天蚕素,是一类具有抗菌能力的小分子多肽,热稳定性好,抗菌谱广,在各个应用领域已经得到广泛的研究。就cecropin抗菌肽结构与功能的关系、作用机制和其在转基因植物抗菌领域中的应用进行综述,阐述了转基因植物表达cecropin抗菌肽具有应用优势和实用价值,cecropin抗菌肽分子结构与作用机制探索的深入能进一步促进转基因植物抗菌研究的发展。

关键词： cecropin 天蚕素作用机制转基因植物

斜纹夜蛾抗菌肽基因Moricin和cecropin时空表达模式及微生物敏感性的分析

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昆虫学报 2012年第9期55卷 1008-1013页

作者：许小霞王爽张文庆孙强王超金丰良华南农业大学资源环境学院广州510642 中山大学生命科学学院有害生物控制和利用国家重点实验室广州510006

抗菌肽是一类具有广谱抗菌活性的小分子多肽,包括抗细菌、病毒和真菌。本研究以斜纹夜蛾Spodoptera litura(Fabricius)为研究对象,利用Real-timePCR和半定量RT-PCR检测了其体内两个重要抗菌肽基因cecropin和Moricin的时空表达模式及其... 详细信息

抗菌肽是一类具有广谱抗菌活性的小分子多肽,包括抗细菌、病毒和真菌。本研究以斜纹夜蛾Spodoptera litura(Fabricius)为研究对象,利用Real-timePCR和半定量RT-PCR检测了其体内两个重要抗菌肽基因cecropin和Moricin的时空表达模式及其对微生物的敏感性。Real-timePCR检测结果表明,抗菌肽基因cecropin和Moricin在斜纹夜蛾各个龄期(1,2,3,4,5和6龄幼虫、蛹、成虫)都有表达,随着龄期的增长,表达量逐渐增加,6龄幼虫中表达量达到最高。半定量RT-PCR检测表明,抗菌肽基因cecropin和Moricin在7个不同组织(气管、卵巢、马氏管、体壁、中肠、脂肪体和血细胞)中都有表达,cecropin在脂肪体中表达量最高,而Moricin在血细胞中表达量相对较高;RT-PCR检测表明,cecropin和Moricin的表达在大肠杆菌Escherichia coli诱导后迅速上升,其中诱导后8h达到高峰期,一直持续到48h。不同病原微生物(大肠杆菌***、金黄色葡萄球菌Staphyloccocus aureus和白僵菌Beauveria bassiana)分别诱导后,Real-timePCR检测表明,斜纹夜蛾抗菌肽基因cecropin和Moricin对大肠杆菌的刺激最为敏感。本研究结果为进一步研究斜纹夜蛾抗菌肽基因cecropin和Moricin的功能奠定基础。

关键词：斜纹夜蛾抗菌肽 Moricin cecropin 时空表达模式微生物敏感性