莱茵衣藻血红素加氧酶1过表达载体的构建和遗传转化.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 2010 2 1 1 1 2 1 王五姐 魏渊源 刘兆普 杨志敏 1 2 ( , 210095; , 210095) : 莱茵衣藻(Chlamydomonas reinhardti)是一种 3套基因组都能进行遗传转化 真核生物, 作为 一种模式生物, 它 被用于生物学研究 各 个领域目前, 发现血红素加氧酶具有多种功能活性,但关于它 作用机理还不是很清楚本研究利 用分子生物学技术, 构建了莱茵衣藻 HO1过表达载体, 用 Spe I和Bgl II双酶切, DNA 测序, GUS染色, PCR 检测证明表达载 体构建成功将此构建通过农杆菌介导法导入莱茵衣藻细胞中, 获得了能够稳定遗传 转化子上述结果为后续进 一步 功能研究奠定基础 : 血红素加氧酶1(HO1) 莱茵衣藻 遗传转化 农杆菌 Expression Vector Construction ofHeme Oxygenase1 from Chlamydom onas reinhardtii and Transformation 1 1 2 1 W angWujie W ei uanyuan Liu Zhaopu ang Zhim in 1 ( College of LfieScience, NanjingAgricultural University, Nanj ing 210095; 2 The ey Laboratory of MarineBiology, Jiangsu Province, NA U, Nanjing 210095) A bstrac:t The green algaChlamydomonas reinhardtii, a unicellular eukaryote has three genomesIt is a valuablemodelsystem for biological studyRecently, heme oxygenase1(HO1) is shown to playmultiple roles in biologicalprocesses, but the accurate functions of its gene are largely unknownIn this study, we constructed an expression vector harboring HO1 from Chlamydomonas rein hardtiiD igestion withSpe I andBgl II, DNA sequencing, GUS staining and PCR detecting proved that theHO1 gene has been cloned into the expression vectorThe constructswere then transferred into the cells byAgrobacteriumm