摘要
[Objectives] This study was conducted to investigate the function of Dunaliella salina calmodulin kinase(CaM K) gene.[Methods] The sense and antisense gene fragments(223 bp) and spacer sequence(129 bp) of D.salina calmodulin kinase gene were cloned and inserted into the downstream part of the35 S promoter of the eukaryotic expression vector pM DCMGN-Cat.The siRNA expression system of CaM K gene was successfully constructed.The p CaM K-RNAi expression vector was transformed into D.salina cells by the LiA c/PEG-mediated method,giving transgenic D.salina.The expression of CaM K gene was then analyzed by real-time fluorescence quantitative PCR.[Results]The expression of CaM K gene in the transgenic D.salina was significantly reduced,by 70% compared with the control group,suggesting that the expression of CaM K gene was significantly inhibited.The examination of the growth status of D.salina showed that D.salina cell division and proliferation were also affected.It is proved that CaM K gene has a positive regulation effect on the division and proliferation of D.salina cells.[Conclusions] The study provides important information for further elucidating the function and action mechanism of D.salina calmodulin kinase gene.
[Objectives] This study was conducted to investigate the function of Dunaliella salina calmodulin kinase( CaM K) gene. [Methods] The sense and antisense gene fragments( 223 bp) and spacer sequence( 129 bp) of D. salina calmodulin kinase gene were cloned and inserted into the downstream part of the35 S promoter of the eukaryotic expression vector pM DCMGN-Cat. The siRNA expression system of CaM K gene was successfully constructed. The p CaM K-RNAi expression vector was transformed into D. salina cells by the LiA c/PEG-mediated method,giving transgenic D. salina. The expression of CaM K gene was then analyzed by real-time fluorescence quantitative PCR. [Results]The expression of CaM K gene in the transgenic D. salina was significantly reduced,by 70% compared with the control group,suggesting that the expression of CaM K gene was significantly inhibited. The examination of the growth status of D. salina showed that D.salina cell division and proliferation were also affected. It is proved that CaM K gene has a positive regulation effect on the division and proliferation of D. salina cells. [Conclusions] The study provides important information for further elucidating the function and action mechanism of D. salina calmodulin kinase gene.
基金
Supported by National Natural Science Foundation of China(31472260,30972240)。
