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层理鞭枝藻藻红蓝蛋白操纵子F基因的克隆和表达 被引量:8

CLONING AND OVEREXPRESSING OF PHYCOERYTHROCYANIN OPERON F-GENE OF MASTIGOCLADUS LAMINOSUS IN ESCHERICHIA COLI
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摘要 以层理鞭枝藻总 DNA为模板 ,用 PCR技术扩增得到藻红蓝蛋白操纵子 F基因(pec F) ,然后克隆于质粒 p Bluescript sk(+)。将 pec F基因亚克隆于表达载体 p GEMD,所形成重组质粒 p GEMD- pec F在 E.coli BL2 1 (DE3 )中获得 1 0 %外源表达蛋白并形成包涵体。这个表达产物的分子量为 2 2 .5 k Da,与按 DNA序列预测的蛋白分子量一致。经藻红蓝蛋白α-亚基的重组实验证明 ,该 pec F基因编码的表达产物 Pec F与藻红蓝蛋白操纵子 E基因(pec E)的表达产物 Pec E共同存在时 ,具有藻红蓝蛋白裂合酶活性。 By PCR method, apo phycoerythrocyanin operon F gene ( pecF ) of Mastigocladus laninosus was amplified from its genomic DNA,and then cloned in pBluescript The pecF gene was subcloned into the expression vector pGEMD,and then transformed into E coli BL21(DE3) After induction, a new protein of molecular weight 22 5kDa existing in inclusion body was overexpressed The expression product was confirmed to possess phycoerythrocyanin lyase activity
出处 《水生生物学报》 CAS CSCD 北大核心 2002年第2期168-174,共7页 Acta Hydrobiologica Sinica
基金 国家自然科学基金 (编号 :39770 1 75 )资助项目 武汉市晨光计划 (编号 :96 5 0 0 1 0 37-2 1 )
关键词 藻红蓝蛋白 操纵子F基因 基因克隆 基因表达 层理鞭枝藻 Phycoerythrocyanin operon F gene Gene clone Gene overexpression
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参考文献9

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  • 1戴汉川,龙良启,丁光.鲤鱼肥胖基因的分子克隆及在大肠杆菌中的表达[J].动物学报,2005,51(1):95-100. 被引量:5
  • 2王红梅,刘嘉莉,杨萍,刘兰英.聚半乳糖醛酸酶(PG2)中色氨酸残基的化学修饰[J].高等学校化学学报,1995,16(2):211-215. 被引量:4
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