家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,Bm NPV)bro(baculovirus repeated ORF)-d基因普遍存在于多种感染鳞翅目昆虫的多角体病毒基因组中,是杆状病毒中一类重复开放阅读框序列。为了解bro-d基因在病毒感染过程中的作用及其与宿主细胞凋亡之间的关系,本研究利用Red重组技术敲除Bm NPV基因组中的bro-d基因,构建基因缺失型病毒bro-d-ko-Bacmid,并将该缺失型病毒转染家蚕Bm N细胞,利用q RT-PCR技术检测bro-d基因的缺失对病毒基因组复制、转录水平以及抑制凋亡蛋白2基因(inhibitor of apoptosis protein 2,iap2)的影响。结果显示,bro-d基因缺失后会导致病毒基因组的复制水平显著下降(P<0.05),同时病毒的早期基因lef-3、晚期基因vp39和极晚期基因p10的转录水平也都显著下降(P<0.05);iap2的转录水平显著下调(P<0.05)。在bro-d基因缺失型病毒的复制水平明显低于野生型病毒的情况下,仍会导致细胞活力显著下降(P<0.05);bro-d基因缺失引起细胞B...
【英文摘要】
bro(baculovirus repeated ORF)-d gene of Bombyx mori nucleopolyhedrovirus(Bm NPV) is a kind of repeated open reading frame sequence in baculovirus and is widespread in a variety polyhedrosis virus genome which specially infects Lepidopteran insect. In order to understand the role of bro- d gene in viral infection and the relationship with host cell apoptosis, the bro-d gene was knocked out by Red recombination system to construct the bro-d-deficiency type virus(bro-d-ko-Bacmid), and was encapsulated by lipos...
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