The development of viral vector for in vivo gene-delivery and its applications

构建了伪狂犬病病毒（PrV）非必需糖蛋白gG（gX）与lacZ的融合基因，并获得了伪狂犬病病毒（PrV）突变株。通过将其注射到SD大鼠的咬肌或胃中，研究了该突变病毒作为跨神经元示踪剂的特性和有效性。注射滴度为1.0* 10^7p.f.u./ ml最适合示踪研究。注射更高滴度的病毒溶液在2天内杀死动物，表明该突变病毒保留了毒力。咬肌注射的三叉神经运动神经均发生病毒感染，但标记神经在三叉神经中脑神经的病例很少。在蓝斑（LC）、中缝大核（RM）、延髓网状结构（IRt）、Edinger-Westphal核（EW）、结节大细胞核（EW）、延髓网状结构（IRt）等核团内均发现二级或三级神经元。(TM)、下丘脑外侧核。(LH)和室旁下丘脑神经（PVH）。将该病毒注入胃内，可标记孤束核、孤束核、孤束核和孤束核的运动背核（DMN）神经元。这些结果表明，该病毒优先感染躯体和内脏运动神经元，并在中枢神经系统的二级或三级神经元中逆行转运。

Fusion gene of non-essential glycoprotein gG (gX) and lacZ was constructed and pseudorabies virus (PrV) mutants were produced. Characteristics and usefulness of this mutant virus as a transneuronal tracer was investigated by injecting it into masseter or stomach of SD rats. Injection of PrV solution with titer of 1.0*10^7p.f.u./ml was most suitable for the tracing study. Injection of higher titer of viral solution killed the animals within 2 days, indicating the retained virulence of this mutant virus. Although viral infection occurred in motor trigeminal n.neurous in all cases of masseter injection, labeled neurous in the mesencephalic trigeminal n.were very few. Second-or third-order neurons were identified in the locus coeruleus (LC), raphe magnus (RM), medullary reticular formation (IRt), Edinger-Westphal nucleus (EW), tuberal magnocellular n.(TM), lateral hypothalamic n.(LH) and paraventricular hypothalamic n.(PVH). No anterograde tracing was detected in the CNS.Injection of this virus into stomach labeled neurons in the dorsal motor nucleus of N.X (DMN), n.tractus solitarius, n. ambiguus, A5 and PVH.These findings indicate that this virus preferentially infecct somatic and visceral motoneurons and is retrogradely tranported in second-order or third-order neurons in the CNS.