Functional Analysis of Dunaliella salina Calmodulin Kinase Gene

• 作者: Mingfang WANG Weiwei XU Xiangnan GAO Xiaojie CHAI Yuting CONG Zhenyu XING
• 来源期刊: 农业生物技术:英文版
• 学科: 医学
• 关键词: DUNALIELLA SALINA CAMK RNAi LiAc/PEG-mediated method Real-time FLUORESCENCE quantitative PCR
• 年，卷（期）: 2020,（2）
• 页码范围: 10-13
• 页数: 4页
• 分类号: R73

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.