中国农业期刊集成服务平台

当前位置: 首页 > 文章 > 生长抑素shRNA的慢病毒包装及对生长抑素的抑制作用吉林农业大学学报 2011,33 (3) 327-331

Position: Home > Articles > Packing of the Lentivirus Contained shRNA Targeting Somatostatin and Its Inhibitory Effect Journal of Jilin Agricultural University 2011,33 (3) 327-331

生长抑素shRNA的慢病毒包装及对生长抑素的抑制作用

作者：

卢可;刘松财;王楠;曹锦艳;张永亮;郝林琳

单位：

吉林大学农学部公共教学中心;山东省军区物资采购办公室;吉林大学畜牧兽医学院;中国兽医药品监察所

关键词：

生长抑素;shRNA;慢病毒载体;基因沉默

摘要：

采用慢病毒(LV)载体介导siRNA感染宿主细胞,研究其对内源性生长抑素(Somatostatin,SS)的抑制作用。首先将筛选出的psh2与辅助质粒共转染293T细胞包装获得假病毒颗粒LV-sh2,同时,包装产生LV-sh0和LV-GFP作为阴性对照。超速离心,梯度稀释法测定病毒滴度,结果提示所构建的重组病毒滴度约为6×107ifu/mL。病毒感染BHK-21细胞,通过荧光显微镜观察到了GFP的高效表达,在感染细胞的基因组中PCR检测到了病毒基因,表明病毒感染获得了高效的基因转移和表达效率。荧光定量PCR和RIA检测表明,LV-sh2对内源性SS的mRNA和蛋白质水平均有显著的抑制作用(86.49%和78.80%,P<0.05)。

译名：

Packing of the Lentivirus Contained shRNA Targeting Somatostatin and Its Inhibitory Effect

作者：

LU Ke1, LIU Song-cai2,WANG Nan3, CAO Jin-yan4,ZHANG Yong-liang2,HAO Lin-lin21.Teaching Center of Basic Courses,Department of Agronomy,Jilin University,Changchun 130062,China;2.College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China 3.China Institute of Veterinary Drug Control,Beijing 100081,China;4.Supplies Purchasing Office of Shandong Provincial Military Command,Jinan 250014,China

关键词：

somatostatin;shRNA;Lentiviral vector;gene silencing

摘要：

Somatostatin(SS) acts to inhibit secretion on a diverse array. In our research,BHK-21 cells were infected by a Lentiviral-shRNA(Lv-shRNA) to examine whether the endogenous SS would be down-regulated by shRNA.First,the Lv-sh2 was packed by the co-transfection of the screened psh2 and the other three packing plasmids into 293T cells.Meanwhile,the Lv-sh0 and Lv-GFP were packed as the negative controls.Next,high-titer virus stocks(6×107 ifu/mL) obtained by ulatracentrifugation were achieved routinely measuring by infecting BHK-21 cells in ten-fold serial dilutions.Finally,BHK-21 cells were infected with the concentrated viral stocks,and the high infection and expression efficiency were observed under the fluorescence.The shRNA gene was detected to integrate into the genomic DNA of infected cells by PCR analysis.Significant down-regulation of SS mRNA and protein level were detected by Real-time PCR(reduced by 86.5% relative to the control,P<0.05) and RIA technique(lower 75.4% than the control,P <0.05).The results may lay a basis on the studies of SS functions in the cell.

相似文章

以减毒沙门菌为载体的SS和CST双表达DNA疫苗经不同途径免疫犊牛的效果评价 [袁婧, 张欣欣, 周迪, 杨帅, 周家彦, 梁爱心, 杨利国] 畜牧兽医学报 2022 (6) 1914-1924
一种有待开发的饲料添加剂——半胱胺 [苏丽娟, 王艳玲, 阴正兴, 厉素霞] 饲料博览 2002 (8) 47-48
二甲酸钾对断奶仔猪胃功能的影响 [夏双双, 姚文, 邹冰洁, 字正浩, 陆扬, 夏东] 畜牧与兽医 2015,47 (3) 52-55
生长抑素基因工程活载体疫苗——激生1号对波杂山羊增重的影响 [肖慎华, 王子玉, 王锋] 畜牧与兽医 2007,39 (10) 33-34
激生Ⅰ号疫苗免疫育肥猪低营养水平饲养增重试验 [孙元麟, 王寿宽, 赵中兴, 杜念兴] 畜牧与兽医 2001,33 (2) 24-25
半胱胺对生长期仔猪增重及有关激素的影响 [魏智清, 张振汉, 于洪川] 甘肃畜牧兽医 2005,35 (1) 12-13
利用生长轴各激素调控动物生长 [王开云, 张国生, 曾检华, 舒邓群] 江西畜牧兽医杂志 2008 (6) 4-6
生长抑素抑制剂半胱胺对动物生产性能的影响 [彭志猛, 宋立华, 舒邓群, 胡军应] 江西畜牧兽医杂志 2003 (6) 22-24
半胱胺在畜禽生产中的研究应用 [亢守亭, 车向荣] 饲料博览 2004 (6) 43-45
日粮添加半胱胺对淮南麻鸭生长性能和血液中生长抑素水平的影响 [陈敏, 张玲, 曹维维, 李建柱, 黄立, 何敏] 饲料博览 2013 (12) 55-58

导出引用

计量

文章访问数: 14

HTML全文浏览量: 0

PDF下载量: 0

生长抑素shRNA的慢病毒包装及对生长抑素的抑制作用

相似文章

所属期刊

吉林农业大学学报

推荐期刊

畜牧兽医学报

中国兽医学报