慢病毒载体介导的IGF2b基因在鲤鱼背部肌肉组织中过表达对鲤鱼生长的影响

The effect of overexpression of IGF2 bgene mediated by lentiviral vector in dorsal muscle of common carp on growth of common carp(Cyprinus carpio L.)

为进一步了解胰岛素样生长因子2b(IGF2b)基因在靶组织过表达对鲤鱼生长的影响,将含IGF2b基因和增强型绿色荧光蛋白(EGFP)基因的慢病毒载体感染鲤鱼背部肌肉组织,注射剂量(每条)分别为10、20、40μL,探讨基因过表达对鲤鱼生长的影响。结果表明:注射48h后,试验组鲤鱼背部肌肉组织可见绿色荧光,4个月后,与对照组相比,试验组体重、体长、体高和体厚均显著提高。为检测慢病毒处理对鲤鱼血清指标的影响,检测与肝功能、肾功能及IGF2b基因路径相关的指标,结果表明:各组间血清IGF2、IGF1R、IGF2R浓度差异均不显著;与对照组相比,10μL注射组碱性磷酸酶、丙氨酸转移酶、总蛋白、尿素氮显著增高,而20、40μL注射组各血清指标与对照组差异均不显著。为验证试验组目的基因的过表达,通过荧光定量PCR检测试验组及对照组间IGF2b基因的表达情况,结果表明:IGF2b基因在试验鲤鱼背部肌肉组织中成功过表达。正是IGF2b基因在鲤鱼背部肌肉组织中的过表达,促进了鲤鱼的生长,且在试验剂量下病毒液对鲤鱼血清无影响。

In order to explore the effect of overexpression of insulin-like growth factor 2b（IGF2b）gene in target tissue on the growth of common carp,the present research focused on the growth of common carp whose dorsal muscle were injected by lentiviral vector containing enhanced green fluorescent protein（EGFP）gene and IGF2 b gene with the dose of10,20 and 40μL.The result showed that green fluorescent was observed in dorsal muscle of common carp after fortyeight hours.Body weight,length,height and thickness of all treated groups except for non-vector injection were significantly higher than control group（P〈0.05）four months later.In order to examine the effects of such vector containing IGF2 b on serum parameters of common carp,the flag indexes of pathway of IGF2 b gene and function of liver and kidney were observed.There was no significant difference among the different groups about the concentration of IGF2,IGF1 R and IGF2 R.But alanine aminotransferase,alkaline phosphatase,total protein and blood urea nitrogen of 10μL treated group were significantly higher than those in control group（P〈0.05）.In order to determine whether the target gene overexpression of treated group were occurred,the relative real time PCR was used to examine the abundance of IGF2 b gene,the result showed that IGF2 b gene was successfully overexpressed in dorsal muscle of experiment groups.These illustrated that overexpression of IGF2 b gene in dorsal muscle of common carp can improve their growth rate with no significant influence on serum functional parameters of common carp.