The limited viability of mesenchymal stem cells restricts its application in treatment in vivo. Therefore, it is very important to find an experiment method to prolong the viability of mesenchymal stem cells so as to lay foundation for experiment in vivo.OBJECTIVE:
To construct the lentiviral vector encoding survivin gene of mice with clone vector pLV.Des3d.P/hygro.METHODS:
Oligo nucleic acid software was used to design a couple of primers according to survivin gene sequence of mice which is publicated on Genbank. The attB recombination site was added to the two ends of the couple of primers, and mSurvivin and IRES/EmGFP were amplified by PCR. The PCR product after BP reaction was transferred to Stb13, and the correct pDown-mSurvivin and pTail-IRES/EmGFP were selected and sequenced. The mixture of pDown-mSurvivin, pTail-IRES/EmGFP and pLV.Des3d.P/hygro was used to do LR reaction. The lentiviral vector pLV.EX3d.P/hygro-EF1A>mSurvivin>IRES/EmGFP was amplified and sequenced again.RESULTS AND CONCLUSION:
The results of PCR and sequencing showed that the lentiviral vector encoding survivin gene of mice was constructed successfully, which lays foundation for the further study of survivin anti-apoptosis in mesenchymal stem cells.