生物学杂志

生物学杂志 ›› 2025, Vol. 42 ›› Issue (2): 52-.doi: 10.3969/j.issn.2095-1736.2025.02.052

• 研究报告 • 上一篇    下一篇

节旋藻同源重组载体的构建及自然转化研究

王　康1,2， 崔红利1， 李文军1， 秦　松1   

  1. 1. 中国科学院烟台海岸带研究所 海岸带生物学与资源利用研究室， 烟台 264003；
    2. 中国科学院大学， 北京 100049
  • 出版日期:2025-04-18 发布日期:2025-04-14
  • 通讯作者: 秦松，博士，研究员，研究方向为分子藻类学及产品工程，E-mail:sqin@yic.ac.cn
  • 作者简介:王康，研究生，研究方向为微藻基因工程，E-mail:wangkang211@mails.ucas.ac.cn
  • 基金资助:
    国家自然科学基金项目（E411011501）； 2023年度吉林省与中国科学院科技合作高新技术产业化专项项目（2023SYHZ0040）

Construction of homologous recombinant vectors and natural transformation of Arthrospira

WANG Kang1,2, CUI Hongli1, LI Wenjun1, QIN Song1   

  1. 1. Research Laboratory of Coastal Zone Biology and Resource Utilization, Yantai Institute of Coastal Zone Research,
    Chinese Academy of Sciences, Yantai 264003; 2. University of Chinese Academy of Sciences,
    Beijing 100049, China
  • Online:2025-04-18 Published:2025-04-14

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摘要： 节旋藻（Arthrospira）是一类具有较高经济价值的多细胞丝状蓝藻。然而，节旋藻的遗传改造极为困难，特别是缺乏易于操作的遗传转化技术。为了进一步开发节旋藻基因工程应用潜力，从12株不同的节旋藻中筛选具备自然转化能力藻株并评估同源侧翼序列和插入片段长度对自然转化效率的影响。结果显示，在12株节旋藻中仅S17和S39可通过自然转化实现遗传操作。同源侧翼序列与插入DNA片段长度均影响转化效率，当同源侧翼序列为1500 bp以上时可显著提升阳性率；相反，插入片段长度则与阳性率呈负相关，因此，当插入DNA较长时需要考虑增加同源侧翼序列长度以提高转化效率。此外，研究还表明节旋藻的同质化与持续筛选时间有关，在筛选120 d后同质化藻株可占阳性藻株的50%以上；同质化藻株可在至少6个月内保持遗传稳定。研究结果可为节旋藻作为高价值化合物生产的生物反应器的应用开发提供技术支持。

关键词: 节旋藻, 自然转化, 同源重组, 基因工程, 载体

Abstract: Arthrospirais a multicellular filamentous cyanobacterium with high economic value. However, genetic manipulation ofArthrospirahas been a major challenge, especially the lack of an easy-to-operate genetic transformation technology. To further develop the potential ofArthrospirafor genetic engineering applications, this study attempted to screen strains with natural transformation ability from 12 differentArthrospiraand evaluated the impact of homologous flanking sequence and insertion fragment lengths on natural transformation efficiency. The results showed that only strains S17 and S39 could be genetically modified through natural transformation. Both homologous flanking sequences and insertion DNA fragment lengths influenced transformation efficiency. It was found that increasing the homologous flanking sequences to over 1 500 bp could significantly increase the positive rate. Conversely, the length of the inserted DNA fragment was negatively correlated with the positive rate, so it was necessary to consider increasing the length of homologous flanking sequences when using longer inserted DNA fragments. In addition, this study also confirmed that the homogeneity ofArthrospirawas related to the duration of continuous screening, where after 120 days of screening, homogenous strains accounted for over 50% of the positive strains; these homogenous strains remained genetically stable for at least 6 months. It was expected that this work would provide technical support for the development ofArthrospiraas a bioreactor for producing high-value compounds.

Key words: Arthrospira, natural transformation, homologous recombinant, genetic engineering, vector

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