摘要
[ Objective] This study was to construct an expression vector capable of excising selectable marker gene, further eliminating the effect of marker gene on the functional study of target gene. [ Method] By using Cre/LoxP site-specific recombination system, DsRed2-1 vector was modified by introducing LoxP se- quence and multiple cloning site sequence, then the TK gene was ligated into this vector for negative selection. [ Results] The fragments introduced were recom- bined at the molecular level under induced condition, and the specific red fluorescence was also observed in the recombinant vector transfected cells at the cellular level. [ Conclusion] It is feasible to use this Cre/loxP system to excise marker genes, showing a broad application prospect.
[ Objective] This study was to construct an expression vector capable of excising selectable marker gene, further eliminating the effect of marker gene on the functional study of target gene. [ Method] By using Cre/LoxP site-specific recombination system, DsRed2-1 vector was modified by introducing LoxP se- quence and multiple cloning site sequence, then the TK gene was ligated into this vector for negative selection. [ Results] The fragments introduced were recom- bined at the molecular level under induced condition, and the specific red fluorescence was also observed in the recombinant vector transfected cells at the cellular level. [ Conclusion] It is feasible to use this Cre/loxP system to excise marker genes, showing a broad application prospect.
基金
Supported by Doctoral Funds of Xinjiang Production and Construction Corps(2011BB014)
Guidance Program of Xinjiang Academy of Agricultural Reclamation Sciences(YYD201110)