全长家蝇抗菌肽基因diptericin的克隆
及生物信息学分析

中国媒介生物学及控制杂志 ›› 2009, Vol. 20 ›› Issue (5): 393-396.

全长家蝇抗菌肽基因diptericin的克隆
及生物信息学分析

杨小蓉1，2，金小宝1，曾爱华3，朱家勇1

1 广东药学院基础学院病原生物研究所（广州 510006）； 2 广东药学院附属第一医院检验科（广州 510080）； 3 广东药学院药科学院

收稿日期:2009-06-05 出版日期:2009-10-20 发布日期:2009-10-20
通讯作者: 朱家勇， E?mail: zhujy@gdpu.edu.cn
作者简介:杨小蓉（1972-），女，医学硕士，从事医学昆虫学方面科研工作。E?mail: xiaorongy3196@qq.com
基金资助:
国家自然科学基金（30672670）

Clone of full?length diptericin from Musca domestica and its bioinformatics analysis

1 Pathogenic Organism Institute, Foundation College of Guangdong Pharmaceutical University, Guangzhou 510006, Guangdong Province, China; 2 The First Affiliated Hospital Laboratory of Guangdong Pharmaceutical University, Guangzhou 510080, Guangdong Province, China

Received:2009-06-05 Online:2009-10-20 Published:2009-10-20
Contact: ZHU Jia?yong, E?mail: zhujy@gdpu.edu.cn

摘要/Abstract

摘要：

【摘要】　目的　克隆新的家蝇抗菌肽基因双翅肽（diptericin）的全长序列并对其进行相关生物信息学分析。方法　根据本实验室已经克隆的家蝇抗菌肽基因diptericin片段设计1对引物，运用 cDNA末端快速扩增技术（GeneRacer）、T?A克隆和序列测定，将所得序列与GenBank中已知的diptericin基因进行同源性分析。结果　成功地从免疫诱导的3龄幼虫中得到一个全长433 bp的基因cDNA ，该全长cDNA具有完整的编码框，编码99个氨基酸，相关生物信息学分析结果表明，其与厩蜇蝇diptericinA基因的同源性最高，为77%。结论　初步推断此全长cDNA是家蝇抗菌肽的一个新基因，可能广泛存在于蝇类昆虫中，为进一步研究其生物活性奠定基础。

关键词: 家蝇抗菌肽, diptericin基因, 快速扩增技术, 生物信息学分析

Abstract:

【Abstract】　Objective　To clone the full?length of diptericin gene from Musca domestica larvae and to analyze by bioinformatics. Methods　A pair of primers were synthesized according to the known sequences of diptericin. Its full?length was amplified by GeneRacer rapid amplification of cDNA ends. PCR product was purified and sequenced by T?A cloning and sequence analysis. The sequence amplified in this study had the homology with the sequence registered in GenBank. Results　There were 433 bp nucleotides in the full?length of cDNA, which had complete coding frame and could code 99 amino acids. The bioinformatics analysis showed that it had higher homology compared to diptericin from Stomoxys calcitrans, and the homology was 77%. Conclusion　The full length of cDNA maybe a new gene of antibacterial peptide from M.domestica, widely existed in Musca. It will provide a basis for the further study of its bioactivity.

Key words: Musca domestica, diptericin gene, GeneRacer, Bioinformatics analysis

中图分类号:

R318

杨小蓉1，2，金小宝1，曾爱华3，朱家勇1. 全长家蝇抗菌肽基因diptericin的克隆
及生物信息学分析[J]. 中国媒介生物学及控制杂志, 2009, 20(5): 393-396.

YANG Xiao-rong, JIN Xiao-bao, ZENG Ai-hua, ZHU Jia-yong. Clone of full?length diptericin from Musca domestica and its bioinformatics analysis[J]. Chines Journal of Vector Biology and Control, 2009, 20(5): 393-396.

参考文献

［1］ Rahuma N，Ghenghesh KS，Ben Aissa R，et al. Carriage by the housefly（Musca domestica）of multiple antibiotic?resistant bacteria that are potentially pathogenic to humans，in hospital and other urban environments in Misurata，Libya［J］. Ann Trop Med Parasitol，2005，99（8）：795-802.
［2］ Boman HG，Nilsson?Faye?I，Paul K，et al. Insect immunity. Characteristics of an inducible cell?free antibacterial reaction in hemolymph of Samia cynthia pupae［J］. Infect Immun，1974，10（6）：136-145.
［3］ Irving P， Troxler L， Heuer TS， et al. A genome?wide analysis of immune responses in Drosophila［J］. Proc Natl Acad Sci USA， 2001，98（26）：15119-15124.
［4］ Zambon RA， Nandakumar M， Vakharia VN， et al. The toll path?way is important foran antiviral response in Drosophila［J］. Proc Natl Acad Sci USA，2005，102（20）：7257-7262.
［5］ Hou LX， Shi YH， Zhai P， et al. Antibacterial activity and in vitro anti?tumor activity of the extract of the larvae of the housefly （Musca domestica）［J］. Ethnopharmacology，2007，（111）：227-231.
［6］ Hou LX， Shi YH， Zhai P，et al. Inhibition of foodborne pathogens by Hf?1， a novel antibacterial peptide from the larvae of the housefly （Musca domestica） in medium and orange juice［J］. Food control，2007，（18）：1350-1357.
［7］程璟侠，于娟，赵瑞君，等. 家蝇蛹抗弓形虫抗菌肽的分离纯化［J］. 中国媒介生物学及控制杂志，2008，19（4）：309-311.
［8］ Otvos L Jr. The short proline?rich antibacterial peptide family［J］. Cell Mol Life Sci，2002，59（7）：1138-1150.
［9］ Diffenbach CW， Dveksler GS. PCR Primer： A Laboratory Manual［M］. New York： Cold Spring Harbor Laboratory Press，1995： 268-288.

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