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首页> 外文期刊>Cellular and molecular life sciences: CMLS >Molecular characterization and expression of the antimicrobial peptide defensin from the housefly (Musca domestica)
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Molecular characterization and expression of the antimicrobial peptide defensin from the housefly (Musca domestica)

机译：家蝇抗菌肽防御素的分子表征和表达

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摘要

A 430-bp cDNA encoding the insect antimicrobial peptide defensin was cloned from the housefly, and designated Musca domestica defensin (Mdde). The open reading frame of the cDNA encoded a 92-amino acid peptide with an N-terminal signal sequence followed by a propeptide that is processed by cleavage to a 40-amino acid mature peptide. Northern analysis and in situ hybridization identified the corresponding mRNA in the fat body of bacterially challenged houseflies and in the epidermis of the body wall of naive and challenged house-flies. The Gram-negative bacterium (Escherichia coli) is a strong inducer of the gene. By RT-PCR, Mdde mRNA was also detected in naive and challenged insects. These findings suggest that the defensin gene is constitutively expressed in the epidermis of the housefly body wall. The predicted mature form of Mdde was expressed as a recombinant peptide in E. coli and Pichia pastoris. The recombinant Mdde expressed in Pichia was active against Gram-positive and some Gram-negative bacteria.
机译：从家蝇中克隆了编码昆虫抗菌肽防御素的430 bp cDNA，命名为家蝇防御素（Mdde）。 cDNA的开放阅读框编码具有N端信号序列的92个氨基酸的肽，其后是通过切割为40个氨基酸的成熟肽而加工的前肽。 Northern分析和原位杂交鉴定了在细菌攻击的家蝇的脂肪体内以及幼稚的和攻击的家蝇的体壁表皮中的相应mRNA。革兰氏阴性细菌（大肠杆菌）是该基因的强诱导剂。通过RT-PCR，还可以在幼稚和受攻击的昆虫中检测到Mdde mRNA。这些发现表明防御素基因在家蝇体壁的表皮中组成性表达。 Mdde的预测成熟形式在大肠杆菌和巴斯德毕赤酵母中表达为重组肽。在毕赤酵母中表达的重组Mdde对革兰氏阳性细菌和某些革兰氏阴性细菌具有活性。

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