期刊论文

中文

畜牧兽医学报

0366-6964

2013

44

6

985-992

10.11843/j.issn.0366-6964.2013.06.023

转基因生物新品种培育重大专项（2011ZX08006-002） 广西家畜遗传改良重点实验室培育基地开放课题（桂牧研科201227） 国家自然科学基金（31171192）

本校为第一机构

旨在初步鉴定miR-21靶向肌肉发育相关的基因。通过靶标预测网站发现结节性硬化症蛋白1基因(TSC1)和蛋白磷酸酶3催化亚基-α-亚型基因(PPP3CA)基因的3'非编码区(3'UTR)均具有与miR-21结合的潜在靶位点;以通城猪为试验材料,扩增其TSC1和PPP3CA基因的3'UTR,经NotⅠ和XhoⅠ双酶切后回收目的片段,连接到双荧光素酶载体psiCHECK中;将重组载体pCHE-TSC1-3'UTR和pCHE-PPP3CA-3'UTR分别转染到猪髋动脉内皮细胞(PIEC)和猪肾细胞(PK15)中,检测其荧光素酶活性。结果显示,当2个靶标载体分别与miR-21共转时,miR-21过表达组在PIEC细胞和PK15细胞中的荧光素酶活性均显著低于对照组(P>0.05)。研究结果表明,猪miR-...

The study was to investigate the regulation mechanism of miR-21 in skeletal muscle de- velopment. Using computational target prediction programs, we identified TSC1 and PPP3CA genes as the potential targets of miR-21 which can specify bind to their 3/ UTR regions. The 3r UTR regions of these two genes were amplified by PCR from Tongcheng pigs genomic DNA. The PCR productions were digested by double digestion with Not I and Xho I. We got the pCHE- TSC1-31UTR and pCHE-PPP3CA-31UTR vectors after digested products were ligated into the vector psiCH...