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首页> 外文期刊>BMC Biotechnology >Increase in the astaxanthin synthase gene (crtS) dose by in vivo DNA fragment assembly in Xanthophyllomyces dendrorhous
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Increase in the astaxanthin synthase gene (crtS) dose by in vivo DNA fragment assembly in Xanthophyllomyces dendrorhous

机译：树突状黄单胞菌体内DNA片段组装增加虾青素合酶基因（crtS）剂量

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Background Xanthophyllomyces dendrorhous is a basidiomycetous yeast that is relevant to biotechnology, as it can synthesize the carotenoid astaxanthin. However, the astaxanthin levels produced by wild-type strains are low. Although different approaches for promoting increased astaxanthin production have been attempted, no commercially competitive results have been obtained thus far. A promising alternative to facilitate the production of carotenoids in this yeast involves the use of genetic modification. However, a major limitation is the few available molecular tools to manipulate X. dendrorhous . Results In this work, the DNA assembler methodology that was previously described in Saccharomyces cerevisiae was successfully applied to assemble DNA fragments in vivo and integrate these fragments into the genome of X. dendrorhous by homologous recombination in only one transformation event. Using this method, the gene encoding astaxanthin synthase ( crtS ) was overexpressed in X. dendrorhous and a higher level of astaxanthin was produced. Conclusions This methodology could be used to easily and rapidly overexpress individual genes or combinations of genes simultaneously in X. dendrorhous , eliminating numerous steps involved in conventional cloning methods.
机译：背景技术黄单胞菌是一种与生物技术有关的担子菌酵母，因为它可以合成类胡萝卜素虾青素。但是，野生型菌株产生的虾青素水平较低。尽管已经尝试了促进虾青素产量增加的不同方法，但迄今为止尚未获得商业竞争性结果。一种促进该酵母中类胡萝卜素生产的有前途的替代方法涉及基因修饰的使用。但是，一个主要的限制是很少有可用的分子工具来操纵X. dendrorhous。结果在这项工作中，先前在酿酒酵母中描述的DNA组装方法已成功应用于体内组装DNA片段，并且仅在一个转化事件中通过同源重组将这些片段整合到X. dendrorhous的基因组中。使用这种方法，编码虾青素合酶（crtS）的基因在X. dendrorhous中过表达，并产生更高水平的虾青素。结论该方法可用于轻松快速地同时在X. dendrorhous中过表达单个基因或基因组合，从而消除了常规克隆方法涉及的许多步骤。

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