沈阳农业大学学报2016，Vol.47Issue(5)：559-566,8.
慢病毒介导EGFP在小鼠胚胎的体外表达
Expression of EGFP in Mouse Embryos Mediated by Lentivirus
摘要
Abstract
In this research, transparent injection, cytoplasm injection and transparent incision with lentivirus co-culture method were used to infect 1-cell stage mouse embryos, and to explore the different developmental stages of embryos infected with EGFP gene expression in the three methods. EGPF gene was extracted from the above three methods of lentivirus infection in different developmental stages of embryos. With EGFP as a template, a 630bp segment of EGFP gene was amplified. Different developmental stages of embryos were transfected by lentivirus, and the amplified EGFP fragment (630bp) were obtained. When the virus was injected below transparent film, the EGFP gene positive rate of 90pL injection group (group C) and 60pL injection group (group B) was significantly better than that of 30pL injection group (group A) (p<0.05). When the virus was injected to the cytoplasm, group C was significantly better than the other group (p<0.05), but later in group A and B difference of embryonic development was not significant (p>0.05). When the embryos with transparent incision were cultured with different concentrations of lentivirus, from 1-cell stage to 4-cell stage embryos among the groups were not significantly differnent (p>0.05), while later the 2×103 cfu•μL-1 group and 3×103 cfu•μL-1 group were significantly better than 1×103 cfu•μL-1 group(p<0.01), but with no difference between 2× 103 cfu•μL-1 group and 3×103cfu•μL-1 group (p>0.05). Transparent film or cytoplasm injection with the lentivirus, 90pL (viral titer of 3×103 cfu•μL-1) injection group gave the best effect expression in vitro of EGFP gene; the embryos with transparent incision were cultured with different concentrations of lentivirus, 3×103 cfu•μL-1 titer group showed the best effect expression in vitro of EGFP gene.关键词
慢病毒/胚胎/EGFP/体外表达Key words
lentivirus/embryo/EGFP/expression in vitro分类
生物科学引用本文复制引用
许诺,贾俊龙,王维,朱玉博,王雪晗,白文林,罗光彬..慢病毒介导EGFP在小鼠胚胎的体外表达[J].沈阳农业大学学报,2016,47(5):559-566,8.基金项目
农业部转基因生物新品种培育重大专项基金项目(H022020060420) （H022020060420）
