[Application of microarrays in screening the antibacterial peptide associated genes of Musca domestica].
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.
Liu
Leishan
L
4 Department of Clinical Laboratory, Fourth Hospital of Changsha, Changsha 410006, China.
Jin
Xiaobao
X
Zhu
Jiayong
J
Xiao
Ping
P
Li
Yuanxiang
Y
Gong
Jianwu
J
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics*; D001483:Base Sequence; D020869:Gene Expression Profiling; D017344:Genes, Insect; D006793:Houseflies / Q000235:genetics* / Q000254:growth & development; D007814:Larva / Q000235:genetics / Q000254:growth & development; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry
D023181:Antimicrobial Cationic Peptides; D015345:Oligonucleotide Probes