摘要
目的运用原位杂交方法检测家蝇抗菌肽基因dipericin组织表达模式。方法基于前期实时荧光定量PCR的结果 ,选取病原诱导12 h后的家蝇三龄幼虫,提取其RNA,克隆出dipericin保守序列,应用地高辛标记的diptericin基因反义RNA探针,对经多重耐药的大肠杆菌和耐甲氧西林葡萄球菌混合液刺激前后的家蝇幼虫组织切片进行组织原位杂交。结果 diptericin基因在诱导和未诱导家蝇幼虫的体壁和肌肉均未表达,在诱导后的脂肪体及中肠有高表达,在未诱导的家蝇幼虫部分组织内也有检测到阳性信号,如气管有很弱的表达,而马氏管则诱导与未诱导均有表达。结论家蝇抗菌肽基因diptericin在家蝇抵御外界微生物感染免疫防御中,有着极其重要的作用。
Diptericins are effective against gram negative bacterial strains. In this study, digoxin-labeled cRNA probe was synthesized by in vitro transcription and used to detect the spatial expression of the different tissue from the Musca domestica third instar larvae which was unchallenged or challenged with bacterial. We found that in unchallenged tissue of larvae, diptericin was detected in trachea and Malpighian tubules; while in challenged tissue of larvae, diptericin was detected stronger signal in the fat body and midgut as compared with unchallenged larvae. But we was not detected any signal in the body wall and muscles in unchallenged and challenged larvae. All the result revealed that the fat body and midgut of Musca domestica are the first defense line in against microbe, which is a foundation to further explore insect immune function.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2012年第8期679-682,共4页
Immunological Journal
基金
国家自然科学基金(30672670)
