蓝藻Synechococcus sp.PCC 7942穿梭表达质粒的构建及胸腺素α_1基因的表达  

CONSTRUCTION OF SHUTTLE EXPRESSION VECTOR AND EXPRESSION OF THYMOSIN α_1 IN SYNECHOCOCCUS SP.PCC7942

作　　者：章军[1] 秦燕[1] 欧阳青[1] 徐虹[1] 周克夫[1] 刘仁海[1] 楼士林[1] 

机构地区：[1]细胞生物学与肿瘤细胞工程教育部重点实验室厦门大学生命科学学院,厦门361005

出　　处：《实验生物学报》2003年第1期1-4,共4页Acta Biologiae Experimentalis Sinica

基　　金：国家863资助项目(863-819-04-03)

摘　　要：利用本实验室构建的含蓝藻Plectonema boryanum内源小质粒的穿梭质粒pPRS-1,改建成含热诱导启动子、泛素融合的胸腺素α_1(UB-Tα_1)目的基因、卡那霉素抗性选择标记、rbcs终止子的新的穿梭表达重组质粒pPRFUT。将这种重组质粒转化蓝藻Synechococcus sp.PCC7942,通过抗性筛选获得了具卡那霉素抗性的转化藻株。经Southern-blot杂交证实,穿梭表达质粒已转入蓝藻Synechococcus sp. PCC7942细胞中;在42℃热诱导30min后,目的基因UB-Tα_1得到较高水平表达,表达量约占总蛋白量的7.5%。The shutter expression vector pPREUT was constructed from the plasmid pPRS-1 containing the en dogenous snail plasmid of Plectonema boryanum. The expression elements such as heat shock gene groESL promoter, foreign gene Ub-thymosin α1, rbcS polyA terminator and Kanamycin resistance gene were all included. The shutter plasmid pPREUT was direcdy transferred into Synechococcus sp. PCC7942. The transformants were obtained through Kanamycin screening. Southern blotting analysis showed that the shutter plasmid have been transferred into Synechococcus sp. PCC7942. After induction by heat shock(42℃) for 30 min, the foreign protein LIR-Tα1 was ex pressed efficiently, which reacted 7.5% of total amount protein.

关 键 词：蓝藻 穿梭质粒 胸腺素Α1 基因表达 

分 类 号：Q786[生物学—分子生物学]