单纯疱疹病毒Ⅰ型扩增子载体转移脑啡肽基因在神经细胞的表达

目的：研究采用单纯疱疹病毒I型扩增子载体将脑啡肽基因导入神经细胞，观察载体lacZ基因和外源脑啡肽基因的表达。方法：实验共分4组；A组为对照组，未加病毒悬液的神经细胞；B组，加病毒悬液的神经细胞培养2d；C组，加病毒悬液的神经细胞培养3d；D组，加病毒悬液的神经细胞培养至第10d。留部基用放射免疫法检测外源性脑啡肽基因的表达。用X-gal原位检测lacZ基因表达反映扩增子病毒的存在和滴度。结果：L－脑啡肽放射免疫检测证实转基因神经细胞L－脑啡肽含量较对照组明显升高（P＜0．05），X-gal染色证实加入假病毒悬液的神经细胞有30％以上的细胞蓝染.结论：HSV－1扩增子假病毒能将外源脑啡肽基因导入神经细胞，并使之有效表达。

Experimental study on the expression of enkephalin gene transferred into neural cells by Herpes Simplex Virus type I amplicon vector

ve Neurotropic Herpes Simplex Virus type I(HSV-I) amplicon vector was adopted to transfer the enkephalin gene into neural cells. The purpose of the present study was to determine whether the express of LacZ gene in the vector and the transferred enkephalin gene could be effectively detected. Methods The mixed stock of the packaged and amplified recombinant plasmid was used to infect the cultured embryonic forebrain cortex cells. The study consisted of 4 groups: in group A in vitro cultured neural cells without the mixed virus stock served as control; in group B the neural cells were added to the mixed virus stock and cultured for 2 days; in group C and D the neural cells were added to the mixed virus stock as in group B but were cultured for 3 days and 10 days respectively. The medium was collected to detected HPPE expression by radioimmunoassay. The lacZ gene expression was detected by X-gal staining. Results L-enkephalin radioimmunoassay showed that the neural cell containing transferred gene had significantly higher L-enkephalin content as compared with those in control group(P<0.05) .The rate of cells expressing beta-galactosidase was up to 30% . Conclusions The amplicon virus we packaged and amplified has high infective ability and can transfer enkephalin gene into neural cells and have it expressed efficiently.