重庆医学Issue(34)：4157-4159,3.DOI:10.3969/j.issn.1671-8348.2013.34.020
大鼠CGRP基因重组慢病毒载体的构建及滴度测定
Construction and titration of rat CGRP gene recombinant lentivirus
摘要
Abstract
Objective To construct lentiviral vector carrying rat′s calcitonin gene-related peptide(CGRP) gene for the following-up study on the function of CGRP .Methods CGRP gene segment was subcloned into shuttle plasmid ,become Puc57-CGRP .The pLenO-DCE-CGRP expression vector was be constructed by double digests .The pLenO-DCE-CGRP and 4 auxiliary packaging plas-mids were co-transfected into 293T cells .Cells were cultured for 48 hours .The supernant was collected and concentrated ,and then the viral titers were tested by multiple proportions dilution method and flow cytometer .The expression levels of CGRP were detec-ted in CGRP-modified 293T cells by Real-time PCR .Results The results of digestion and sequencing show that the pLenO-DCE-CGRP vector was constructed successfully .The titer of the lentiviral particles was 5 .1 × 108 TU /mL .Conclusion The high-titer lentvirus vector containing CGRP gene is constructed successfully ,which lay a foundation for transfecting mesenchymal stem cell (MSC) and studying the function of CGRP .关键词
降钙素基因相关肽/慢病毒属/DNA ,重组Key words
calcitonin gene-related peptide/lentivirus/DNA ,recombinant引用本文复制引用
陈攀科,石蓓,许官学,刘志江,王冬梅..大鼠CGRP基因重组慢病毒载体的构建及滴度测定[J].重庆医学,2013,(34):4157-4159,3.基金项目
国家自然科学基金资助项目（81060014）。 （）
