Bai Xiaodong, Grewal Parwinder S
Department of Entomology, The Ohio State University-OARDC, 1680 Madison Avenue, Wooster, OH 44691, USA.
Mol Biochem Parasitol. 2007 Dec;156(2):162-6. doi: 10.1016/j.molbiopara.2007.07.018. Epub 2007 Aug 2.
Entomopathogenic nematode Heterorhabditis bacteriophora infective juveniles (IJs) employ multiple strategies to combat with insect innate immune system after invasion. We employed suppressive subtractive hybridization (SSH) technique to study the gene expression in the IJs upon contact with the insect hemolymph in vitro. We identified two clones having higher expression levels in the IJs than IJs treated with insect hemolymph. The differential expression levels were confirmed by Northern blot hybridization with reference to the constitutive expression level of Heterorhabditis bacteriophora actin2 gene. Clone HbGPS11.C1G02 encoded a phosphofructokinase (PFK) with a 2.5 kb transcript and clone HbGPS11.C4C08 corresponded to a 2.1 kb transcript encoding a protein that had weak similarity to trans-sialidase from Trypanosoma cruzi. The differential expression of PFK in H. bacteriophora IJs before and during recovery process represented a switch from active movement in search of insect hosts to a state of combating insect innate immune system. This first report of H. bacteriophora differential gene expression provides a glimpse at the gene expression profile of H. bacteriophora IJ recovery process.
昆虫病原线虫嗜菌异小杆线虫的感染性幼虫(IJs)在侵入昆虫后会采用多种策略来对抗昆虫的先天免疫系统。我们运用抑制性消减杂交(SSH)技术研究了IJs在体外与昆虫血淋巴接触后的基因表达情况。我们鉴定出两个在IJs中表达水平高于用昆虫血淋巴处理过的IJs的克隆。通过参照嗜菌异小杆线虫肌动蛋白2基因的组成型表达水平,Northern印迹杂交证实了差异表达水平。克隆HbGPS11.C1G02编码一种具有2.5 kb转录本的磷酸果糖激酶(PFK),克隆HbGPS11.C4C08对应于一个2.1 kb的转录本,该转录本编码一种与克氏锥虫的转唾液酸酶有微弱相似性的蛋白质。嗜菌异小杆线虫IJs在恢复过程之前和期间PFK的差异表达代表了从主动寻找昆虫宿主的运动状态转变为对抗昆虫先天免疫系统的状态。嗜菌异小杆线虫差异基因表达的这一首次报道让我们得以初步了解嗜菌异小杆线虫IJs恢复过程中的基因表达谱。