Bkt 基因和 crtR-B 基因在集胞藻 PCC 6803 中的 重组表达及功能分析
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国家自然科学基金项目(41876188);山东省自然科学基金项目(ZR2018ZB0210)

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Recombinant Expression and Functional Analysis of bkt Gene and crtR-B Gene in Synechocystis sp. PCC 6803
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National Natural Science Foundation of China (41876188), and Natural Science Foundation of Shandong Province (ZR2018ZB0210)

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    摘要:

    将雨生红球藻中的 β-胡萝卜素酮化酶基因(bkt)和 β-胡萝卜素羟化酶基因(crtR-B)经密码子优 化后,通过自然转化法分别转入集胞藻 PCC 6803 基因组中。高效液相色谱分析显示:转入 bkt 基因的 细胞产生角黄质的同时,海胆酮含量减少;转入 crtR-B 基因的细胞产生金盏花黄质的同时,玉米黄素 含量减少。实验结果表明,外源的 β-胡萝卜素酮化酶基因将海胆酮转化为角黄质,外源的 β-胡萝卜素 羟化酶基因将玉米黄素转化为金盏花黄质。该文利用代谢工程策略,在集胞藻 PCC 6803 中构建类胡萝 卜素生物合成途径,为通过代谢工程在集胞藻 PCC 6803 中生产虾青素奠定了基础。

    Abstract:

    The β-carotene ketolase gene (bkt) and β-carotene hydroxylase gene (crtR-B) from Haematococcus pluvialis were codon-optimized and transferred to Synechocystis sp. PCC 6803 genome by natural transformation method. High performance liquid chromatography analysis showed that cells transfected with bkt gene produced canthaxanthin, while echinone decreased; the cells with crtR-B gene produced adonixanthin, while zeaxanthin was reduced. The results showed that the exogenous β-carotene ketolase converted echinone to canthaxanthin and the exogenous β-carotene hydroxylase converted zeaxanthin into adonixanthin. In this paper, the pathway of astaxanthin biosynthesis in Synechocystis sp. PCC 6803 was constructed by metabolic engineering strategy, which laid a foundation for astaxanthin production in Synechocystis sp. PCC 6803 with metabolic engineering.

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引文格式
刘亚铭,王康,崔玉琳,等. Bkt 基因和 crtR-B 基因在集胞藻 PCC 6803 中的 重组表达及功能分析 [J].集成技术,2021,10(5):96-103

Citing format
LIU Yaming, WANG Kang, CUI Yulin, et al. Recombinant Expression and Functional Analysis of bkt Gene and crtR-B Gene in Synechocystis sp. PCC 6803[J]. Journal of Integration Technology,2021,10(5):96-103

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  • 在线发布日期: 2021-09-15
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