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首页> 外文期刊>Bioprocess and Biosystems Engineering >A transgene expression system for the marine microalgae Aurantiochytrium sp. KRS101 using a mutant allele of the gene encoding ribosomal protein L44 as a selectable transformation marker for cycloheximide resistance
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A transgene expression system for the marine microalgae Aurantiochytrium sp. KRS101 using a mutant allele of the gene encoding ribosomal protein L44 as a selectable transformation marker for cycloheximide resistance

机译:海洋微藻Aurantiochytrium sp。的转基因表达系统。 KRS101使用编码核糖体蛋白L44的基因的突变等位基因作为环己酰亚胺抗性的选择性转化标记

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摘要

In the present study, we established a genetic system for manipulating the oleaginous heterotrophic microalgae Aurantiochytrium sp. KRS101, using cycloheximide resistance as the selectable marker. The gene encoding ribosomal protein L44 (RPL44) of Aurantiochytrium sp. KRS101 was first identified and characterized. Proline 56 was replaced with glutamine, affording cycloheximide resistance to strains encoding the mutant protein. This resistance served as a novel selection marker. The gene encoding the △12-fatty acid desaturase of Mortierella alpina, used as a reporter, was successfully introduced into chromosomal DNA of Aurantiochytrium sp. KRS101 via 18S rDNA-targeted homologous recombination. Enzymatic conversion of oleic acid (C18:1) to linoleic acid (C18:2) was detected in transformants but not in the wild-type strain.
机译:在本研究中,我们建立了一个遗传系统,用于处理油质异养微藻类Aurantiochytrium sp。 KRS101,使用对环己酰亚胺的抗性作为选择标记。编码Aurantiochytrium sp的核糖体蛋白L44(RPL44)的基因。首先确定并表征了KRS101。脯氨酸56被谷氨酰胺替代,从而使环己酰亚胺对编码突变蛋白的菌株具有抗性。该抗性用作新颖的选择标记。成功地将报道了高山被孢霉Mortierella alpina的△12-脂肪酸脱氢酶的基因成功导入到Aurantiochytrium sp的染色体DNA中。 KRS101通过18S rDNA靶向的同源重组。在转化体中检测到油酸(C18:1)酶转化为亚油酸(C18:2),但在野生型菌株中未检测到。

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  • 来源
    《Bioprocess and Biosystems Engineering》 |2013年第9期|1191-1197|共7页
  • 作者单位

    Applied Microbiology Research Center, Bio-Materials Research Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup,Jeonbuk 580-185, Korea;

    Applied Microbiology Research Center, Bio-Materials Research Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup,Jeonbuk 580-185, Korea;

    Applied Microbiology Research Center, Bio-Materials Research Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup,Jeonbuk 580-185, Korea;

    Applied Microbiology Research Center, Bio-Materials Research Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup,Jeonbuk 580-185, Korea;

    Systems and Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 305-333, Korea;

    Department of Chemical and Biomolecular Engineering and Advanced Biomass R&D Center, KAIST, 291 Daehakno,Yuseong-gu, Daejeon 305-701, Korea;

    Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Kobe 657-8501, Japan;

    Applied Microbiology Research Center, Bio-Materials Research Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup,Jeonbuk 580-185, Korea;

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  • 正文语种 eng
  • 中图分类
  • 关键词

    Aurantiochytrium; Ribosomal protein; Cycloheximide resistance; Gene transformation;

    机译:金眼鱼;核糖体蛋白环己酰亚胺抗性;基因转化;

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