一种高效的莱茵衣藻电转化方法  

An efficient electroporation electroporation transformation method for Chlamydomonas reinhardtii

作　　者：刘佳 何炫程 项晨 马伯宁 韦航涛 杨明峰 LIU Jia;HE Xuancheng;XIANG Chen;MA Boning;WEI Hangtao;YANG Mingfeng(Key Laboratory of Urban Agriculture (North China) Ministry of Agriculture,College of Biological Science and Engineering,Beijing University of Agriculture,Beijing 102206,China)

机构地区：[1]北京农学院生物科学与工程学院农业部华北都市农业重点实验室,北京102206

出　　处：《北京农学院学报》2019年第2期5-9,共5页Journal of Beijing University of Agriculture

基　　金：国家自然科学基金项目(31370674)

摘　　要：【目的】为了探明莱茵衣藻Chlamydomonas reinhardtii最佳的转化条件。【方法】将融合基因4CL-3aSTS与绿色荧光蛋白(GFP)分别作为目的基因,以aphVIII为抗性基因,采用电转化法将两种目的基因分别转入野生型(WT)莱茵衣藻和细胞壁缺失型莱茵衣藻(CC425)中。【结果】用巴龙霉素抗性筛选以及通过PCR验证与荧光共聚焦显微镜检测,证实目的基因在两种莱茵衣藻中成功表达。【结论】获得了一种高效的莱茵衣藻电转化方法,野生型衣藻转化率为11.07%,细胞壁缺失型衣藻转化率为49.67%。【Objective】To find out the best transformation condition of Chlamydomonas reinhardtii.【Methods】The fusion gene of 4CL - 3a - STS and the reporter gene encoding green fluorescent protein ( GFP ) were used as the target genes,and the aminoglycoside 3'-phosphotransferase type VIII (APHVIII) encoding gene( aphVIII ) was used as the resistance gene.The two target genes were transferred into the Chlamydomonas reinhardtii wild-type (WT) and cell wall-deficient type (CC425) by electric transformation,respectively.【Results】Two target genes were successfully transferred and obviously expressed in Chlamydomonas reinhardtii strains verified by paromomycin resistance screening,PCR and fluorescence confocal microscopy.【Conclusion】An efficient electroporation transformation method of Chlamydomonas reinhardtii was obtained.The transformation rate of wild-type Chlamydomonas was 11.07 % while the cell wall-deficient Chlamydomonas was 49.67 %.

关 键 词：莱茵衣藻 电转化 融合基因 绿色荧光蛋白 巴龙霉素 

分 类 号：Q812[生物学—生物工程]