Construction and identification of lentiviral vector-mediated short hairpin RNA of human EGF-like domain 7 gene
10.13699/j.cnki.1001-6821.2017.19.027
- VernacularTitle:类表皮生长因子域7基因shRNA慢病毒表达载体的构建及鉴定
- Author:
Ai-Yue WANG
1
;
Qiang LI
;
Qiong-Guan XU
;
Da-Yuan LIU
;
Peng-Xiang XU
;
Dan YU
Author Information
1. 海口市人民医院神经内科
- Keywords:
EGF-like domain 7;
RNA interference;
short hairpin RNA;
lentivirus
- From:
The Chinese Journal of Clinical Pharmacology
2017;33(19):1946-1948
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the LV3-human EGF-like domain 7-targeted small hairpin RNA (LV3-hEGFL7-shRNA)-expressing plasmid system.Methods Sequence of hEGFL7-targeted shRNA was designed by Oligo Designer 3.0 based on the mRNA sequence of hEGFL7 which was obtained from the Genbank.They were recombined with the plasmid pGLV3/H1/GFP + Puro,and then those plasmids were identified by gene sequencing to make sure they were correctly connec-ted.Then those plasmids were transfected into 293T cells and fluorescence expression was observed.Results The sequence of EGFL7-shRNA was demonstrated that the inserted sequences were correct,and the titer of virus was 2 × 10s TU · mL-1.Conclusion The hEGFL7-targeted shRNA expressing plasmids were successfully constructed,identification can meet test requirements.
