碳离子束诱变选育高产角鲨烯类酵母及电转化初探  

Mutation and selection of high squalene production yeast Pseudozyma sp.induced by carbon-ions beam irradiation and its electrotransformation

作　　者：肖艳 王璐[2] 王森 丛培虎 陆栋[4] 冯银刚[1] 崔球[1] 宋晓金[1] XIAO Yan;WANG Lu;WANG Sen;CONG Peihu;LU Dong;FENG Yingang;CUI Qiu;SONG Xiaojin(Key Laboratory of Biofuels,Chinese Academy of Sciences/Shandong Provincial Key Laboratory of Synthetic Biology/Shandong Energy Institute/Qingdao Institute of Bioenergy and Bioprocess Technology,Chinese Academy of Sciences,Qingdao 266101,China;SDU-ANU Joint Science College,Shandong University,Weihai 264209,China;Shandong Science and Technology Service Development and Promotion Center,Jinan 250101,China;Institute of Modern Physics,Chinese Academy of Sciences,Lanzhou 730000,China)

机构地区：[1]中国科学院生物燃料重点实验室/山东省合成生物学重点实验室/山东省能源研究院/中国科学院青岛生物能源与过程研究所,山东青岛266101 [2]山东大学澳国立联合理学院,山东威海264209 [3]山东省科技服务发展推进中心,山东济南250101 [4]中国科学院近代物理研究所,甘肃兰州730000

出　　处：《南方水产科学》2022年第2期98-104,共7页South China Fisheries Science

基　　金：国家重点研发计划项目(2019YFD0901904);山东省重点研发计划项目(2019GHY112006)。

摘　　要：为进一步提高角鲨烯产量,以海洋产角鲨烯类酵母(Pseudozyma sp.)SD301为出发菌株,采用碳离子(;C;)束诱变技术选育高产角鲨烯的突变菌株,并对突变菌株的电转化条件进行优化。利用100μmol·L^(-1)过氧化氢(H;O;)作为筛选压力,在12C6+束辐照剂量为120 Gy时,获得了比出发菌株具有更高角鲨烯产量的突变株PS120,该突变株在培养3 d后,角鲨烯产量达到1.33 g·L^(-1),角鲨烯质量(细胞质量)达到41.31 mg·g^(-1)。利用绿色荧光蛋白EGFP作为表征标记,对PS120进行电转化条件优化,酶切、电泳和测序研究结果表明900 V电压条件下可成功将编码EGFP的基因转入到PS120细胞中,通过激光共聚焦显微镜进一步证实EGFP蛋白在该细胞中高水平表达。In order to improve the yield of squalene,using Pseudozyma sp.SD301 as the starting strain,we selected the mutant strain with high squalene yield by carbon-ions(;C;)beam irradiation technology,and optimized the electrotransformation conditions of the mutant strain.We used 100μmol·L^(-1)H;O;as the screening pressure.When the carbon heavy ion beam irradiation dose was 120 Gy,the mutant PS120 was obtained with higher squalene yield than the original strain.After 3 days of culture,the squalene yield of the mutant reached 1.33 g·L^(-1)and the squalene mass reached 41.31 mg·g^(-1).The green fluorescent protein EGFP was used as the characterization marker to optimize the electrotransformation conditions of PS120.The results of enzyme digestion,electrophoresis and sequencing show that the gene encoding EGFP could be successfully transferred into PS120 cells under 900 V voltage.The high-level expression of EGFP protein in PS120 cells was further confirmed by laser confocal microscope.

关 键 词：角鲨烯 类酵母 碳离子束 诱变 电转化 

分 类 号：Q939.9[生物学—微生物学]