全长家蝇抗菌肽基因diptericin的克隆及生物信息学分析

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

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433bp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。

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 eDNA 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 nueleotides in the full-length of eDNA, 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 eDNA 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.