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一种新型抗菌肽Hydramacin-1在毕赤酵母中的重组表达、纯化及其抗菌活性 被引量:1
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作者 孟德梅 石林玥 +4 位作者 李文娟 孙雪晴 郭雅君 贾雪霞 樊振川 《天津科技大学学报》 CAS 2019年第4期22-29,共8页
Hydramacin-1是来源于大乳头水螅(Hydra)的上皮防御组织的具有广谱抗菌特性的新型抗菌肽.本研究构建了外泌型重组酵母表达载体pPICZαA-Hydramacin-1,再将线性化的重组表达载体pPICZαA-Hydramacin-1电转入毕赤酵母宿主菌(Pichia pastor... Hydramacin-1是来源于大乳头水螅(Hydra)的上皮防御组织的具有广谱抗菌特性的新型抗菌肽.本研究构建了外泌型重组酵母表达载体pPICZαA-Hydramacin-1,再将线性化的重组表达载体pPICZαA-Hydramacin-1电转入毕赤酵母宿主菌(Pichia pastoris)GS115中,成功构建了重组毕赤酵母表达菌株.在培养温度为28℃、甲醇体积分数为1.5%、诱导表达时长为120h时,蛋白表达量达到最高,重组抗菌肽Hydramacin-1的表达量可达83mg/L.经阳离子交换柱纯化,每升发酵液可得到约9mg纯度为90%的重组抗菌肽Hydramacin-1.经最小抑菌浓度的测定,制备的重组抗菌肽具有广谱抗菌特性,相比于革兰氏阳性菌,革兰氏阴性菌对此抗菌肽更为敏感.本实验将Hydramacin-1抗菌肽首次成功地在毕赤酵母中高效表达,为以后该抗菌肽的理论研究和应用奠定了基础. 展开更多
关键词 抗菌肽 hydramacin-1 毕赤酵母 抗菌活性
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First molluscan antimicrobial peptide hydramacin in Manila clam:molecular characterization and expression analysis
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作者 Youngdeuk Lee Saranya Revathy K +6 位作者 Sukkyoung Lee Ilson Whang Chulhong Oh Do-Hyung Kang Gee-Wook Shin Jehee Lee Mahanama De Zoysa 《Journal of Coastal Life Medicine》 2014年第6期447-452,共6页
Objective:To investigate molecular characterization and the immune responses of Manila clam hydramacin(Rp-hdmc).Methods:cDNA sequence of hydramacin was isolated from Manila clam transcriptome database.Molecular charac... Objective:To investigate molecular characterization and the immune responses of Manila clam hydramacin(Rp-hdmc).Methods:cDNA sequence of hydramacin was isolated from Manila clam transcriptome database.Molecular characterization of hydramacin cDNA was performed by BLAST and SWISS-MODEL bioinformatics programs.Tissue-specific expression and transcriptional regulation after Vibrio tapetis challenge was done by quantitative real time PCR.Results:Rp-hdmc has 291 bp open reading frame(ORF),encoding 97 amino acids with a mature hydramacin consisting of 77 amino acid residues.In un-challenged clam,Rp-hdmc was constitutively expressed in all tested tissues and the highest expression level was detected in gill.After pathogenic bacteria Vibrio tapetis challenge,Rp-hdmc mRNA was up-regulated in gill and hemocytes.Conclusions:We identified hydramacin cDNA(Rp-hdmc)from mollusk Manila clam that shows the characteristic features of hydramacin sequence.It has eight cysteine residues with four disulfide linkages,three helices and two β-strands in secondary structure.Expression results after V.tapetis challenges suggest that Rp-hdmc is involved in immune response against pathogenic bacteria. 展开更多
关键词 Manila clam Ruditapes philippinarum hydramacin Antimicrobial peptide
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