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[Application of microarrays in screening the antibacterial peptide associated genes of Musca domestica].

Authors
  • Liu, Leishan
  • Jin, Xiaobao
  • Zhu, Jiayong
  • Xiao, Ping
  • Li, Yuanxiang
  • Gong, Jianwu
Type
Published Article
Journal
Sheng wu gong cheng xue bao = Chinese journal of biotechnology
Publication Date
Jul 01, 2008
Volume
24
Issue
7
Pages
1300–1305
Identifiers
PMID: 18837412
Source
Medline
License
Unknown

Abstract

To screen the candidate genes associated with Musca domestica antibacterial peptides using DNA microarray technique, the hybrid probes were designed from the conservative domains of the encoded area of the insect antibacterial peptide genes in GenBank with biology software Designer 2.0, and were synthesized by a chemical process, with the assistance of the automated Gen III Microarray Spotter, those oligo probes were printed on a special ready-made glass, and a cDNA microarray was constructed. The total RNA was extracted from the fat body of Musca domestic third-instar larve induced after 24 hours by Escherichia coli and Staphylococcus aureus, the strands of cDNA were labled with fluoresceine Cy3 using the method of reverse transcription PCR, after prehybridization, hybridization and washing procedure, the results of hybridization were scanned using computer system, and the data were analyzed using the software of MIDAS, fifteen valid hybridization signals were detected through two times of hybridization and scanning (the positive samples as a control were excluded). DNA microarray technique can be successfully applied screen the candidate genes associated with Musca domestica antibacterial peptides, and further provide significant evidence to discover its antibacterial peptide new genes.

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