Construction and Identification for Over-expressing Lentiviral Vector of mSema3A / 中国医科大学学报

ABSTRACT

Objective To construct and identify over?expressing lentiviral vector of mSema3A. Methods Sema3A gene of mice was amplified by PCR,then the gene was inserted into plasmids pDown?mSema3A?IRES/EGFPby Gateway technology. The plasmids pLV/EXPNZ?puro?mSema3A?IRES/EGFP were produced by recombination. After sequencing identification,the vector pLV(Exp)?Puro?CMV?mSema3A?IRES/EGFP was packed and condensed. Finally the recombinant vectors were used to transfect 293T cells to obtain virus pools. Results The recombinant lentiviral vectors were 11 538 bp with EGFP marker,and Sema3Agenes were inserted into the lentiviral vector correctly,indicating the over?expressing vector of Sema3A gene in mice was successfully constructed. Conclusion The over?expressing lentiviral vector of mSema3A was constructed correctly, which lay a foundation of screening of over?expressing strains of such gene in specific cells.