水生生物学报2018，Vol.42Issue(2)：416-420,5.DOI:10.7541/2018.053
湖泊微拟球藻高含油突变株的构建及筛选
CONSTRUCTION OF OIL-RICH MUTANTS IN NANNOCHLOROPSIS LIMNETICA
摘要
Abstract
Nannochloropsis are model species for investigating biofuel production by algae.Many marine Nanno-chloropsis genomes have been sequenced recently and their transformation systems have been established.N.limnetica is the only freshwater species, and its genome and transformation system are not yet available. In this study, a genetic transformation method for N.limnetica is established and a zeocin-resistance gene is introduced into the genome to ob-tain mutant library,then high oil-yield mutants are screened from the library.A vector pPha-T1-TUB that has a sh ble gene derived by Nannochloropsis β-tublin promoter and Phaeodactylum tricornutum fcpA terminator was constructed, and the linearized plasmid was introduced to N.limnetica by electroporation.Transformants were able to grow on 1 μg/mL zeocin. PCR detection indicated that 100% of the selected colonies were positive transformants and all resis-tant colonies contained sh ble gene after half a year.Several mutants with higher triacylglycerols content and biomass were obtained. Among them, K26 and G5 showed lower polyunsaturated fatty acid contents than wild type, which meets the requirements of the European Standard EN 14214 (2004) for biodiesel production. This study provides an effi-cient method to obtain advantageous mutants via generating a library of random resistant gene insertion mutations in N. limnetica.关键词
湖泊微拟球藻/三酰基甘油/电转化/突变株Key words
Nannochloropsis limnetica/Triacylglycerols/Electroporation transformation/Mutants分类
农业科技引用本文复制引用
张玲香,陈怡雯,胡晗华..湖泊微拟球藻高含油突变株的构建及筛选[J].水生生物学报,2018,42(2):416-420,5.基金项目
国家自然科学基金面上项目(41576144)资助[Supported by the National Natural Science Foundation of China(41576144)] （41576144）
