小球藻的沼液驯化、抗生素敏感性分析和选择标记筛选  

Biogas Slurry Acclimatization,Antibiotic Sensitivity Analysis,and Selective Marker Screening of Chlorella

作　　者：张红兵[1] 史秀英 李会宣[1] 李磊 李文涛 ZHANG Hongbing;SHI Xiuying;LI Huixuan;LI Lei;LI Wentao(College of Biology Science and Engineering,Heibei University of Economics and Business,Shijiazhuang 050061,China)

机构地区：[1]河北经贸大学生物科学与工程学院,河北石家庄050061

出　　处：《化学与生物工程》2020年第4期22-27,共6页Chemistry & Bioengineering

基　　金：河北省科技厅重点研发项目(18273612D);河北省高等学校科学技术研究项目(ZD2017230);石家庄市科学技术研究与发展计划项目(191240243A);河北经贸大学校内科研项目(2015KYZ01);2019年度河北经贸大学研究生创新计划项目。

摘　　要：通过平板划线法从河北的部分河流中筛选到5株小球藻,经奶牛养殖废水沼液驯化后得到优势藻株,通过18S rDNA序列分析和系统发育树构建对优势藻株种属进行鉴定,并比较了其对G418、氨苄青霉素和潮霉素的敏感性。结果表明,藻株X-1在奶牛养殖废水沼液中的适应能力最强,对沼液中总氮、总磷和COD的去除能力最强;藻株X-1与Chlorella sp.的相似度最高;小球藻X-1对G418的敏感性最强,G418可作为小球藻X-1基因工程中的选择标记抗生素,最佳浓度为160μg·mL^-1。该研究为小球藻X-1利用G418作为选择标记构建表达载体、转化小球藻、培养无菌藻系提供了实验依据。We screened five strains of Chlorella from some rivers of Hebei Province by a streak plate method,obtained a dominant strain after acclimatization by using dairy farming wastewater biogas slurry,identified its species by 18S rDNA sequence analysis and phylogenetic tree,and compared its sensitivity to G418,Ampicillin,and Hygromycin.The results show that the adaptability of algae strain X-1 is the strongest in the dairy farming wastewater biogas slurry,and its removal abilities of total nitrogen,total phosphorus,and COD are the strongest.The similarity between algae strain X-1 and Chlorella sp.is the highest.The sensitivity of Chlorella X-1 to G418 is the strongest.Thus,G418 can be used as the selective marker antibiotics in the genetic engineering of Chlorella X-1,and the optimal concentration is 160μg·mL^-1.This study provides experimental basis for the construction of expression vector,transformation of Chlorella,and culture of aseptic algal lines by using G418 as a selective marker.

关 键 词：小球藻 沼液驯化 抗生素敏感性分析 抗生素标记 筛选 

分 类 号：Q949.2[生物学—植物学]