基因芯片技术筛选家蝇抗菌肽相关基因  

Application of Microarrays in Screening the Antibacterial Peptide Associated Genes of Musca domestica

作　　者：刘雷山[1] 金小宝[2] 朱家勇[2] 肖平[3] 李沅湘[1] 龚建武[1] 

机构地区：[1]长沙市第四医院检验科中心实验室,长沙410006 [2]广东药学院基础学院病原生物研究室,广州510006 [3]中南大学湘雅二医院肾病研究所,长沙410011

出　　处：《生物工程学报》2008年第7期1300-1305,共6页Chinese Journal of Biotechnology

基　　金：国家自然科学基金项目(No.30671832)资助~~

摘　　要：用生物学软件对GenBank中部分昆虫抗菌肽基因编码区保守域设计探针,用直接点样法将探针点印在特制玻片上构建寡核苷酸(Oligonucleotide,oligo)探针微阵列;提取诱导后24h的家蝇三龄幼虫脂肪体总RNA,逆转录成cDNA并标记上荧光标记物Cy3,与构建的oligo探针微阵列杂交,经洗片、扫描处理后进行数据分析。结果在两次重复实验中均检测到有效杂交信号的基因点有15个(不包括阳性对照基因),为进一步发现其新基因提供了依据。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.

关 键 词：基因芯片 家蝇 抗菌肽基因 昆虫抗菌肽基因 筛选 

分 类 号：Q78[生物学—分子生物学]