西安交通大学学报（医学版）2012，Vol.33Issue(6)：786-789,4.
携带NT4-AcSDKP融合基因重组腺相关病毒载体的构建
Construction of NT4-AcSDKP recombinant adeno-associated virus
何娟娟 1马列婷 1王亚文 1惠凌云 1杨广笑 2王全颖2
作者信息
- 1. 西安交通大学医学院第一附属医院检验科,陕西西安710061
- 2. 西安华广生物工程有限公司,陕西西安710025
- 折叠
摘要
Abstract
Objective To construct and produce the NT4-AcSDKP recombinant adeno-associated virus (AAV) and determinate the titer. Methods With plasmid vector pGEM-T Easy/NT4 as the template, NT4-AcSDK was obtained by polymerase chain reaction (PCR) and inserted into vector plasmid PSSHG-CMV to construct the recombinant plasmid pSSCMV-NT4-AcSDKP. The recombinant AAV-NT4-AcSDKP was packaged through co-transfecting 80% of confluent 293 cells with adenovirus full genome plasmid pFG140, helper plasmid pAAV/Ad and the recombinant plasmid. The recombinant AAV-NT4-AcSDKP was titrated by RT-PCR. Results The gene of NT4-AcSDKP was synthesized and the recombinant plasmid pSSCMV-NT4-AcSDKP was constructed successfully. RT-PCR results showed that we had obtained the rAAV of high titer (3. 40 × 1010 copies/mL). Conclusion We have successfully obtained the AcSDKP-expressing rAAV and paved the way for further studies on anti-inflammation, anti-fibrosis, and organ repair.关键词
NT4-AcSDKP/PCR技术/重组腺相关病毒Key words
NT4-AcSDKP/polymerase chain reaction (PCR) technique/recombinant adeno-associated virus (rAAV)分类
生物科学引用本文复制引用
何娟娟,马列婷,王亚文,惠凌云,杨广笑,王全颖..携带NT4-AcSDKP融合基因重组腺相关病毒载体的构建[J].西安交通大学学报（医学版）,2012,33(6):786-789,4.
