As a new expression system, Dunaliella salina (D. salina) has bright prospects and applications in various fields. However, its application is currently restricted because of the low expression and instability of foreign gene in D. salina cells. During genetic operation, transformation is a crucial step for genes expression in D. salina system. Although several transformation methods are existing currently, many inherent deficiencies and limitations still can be found in actual practice. Thus, we attempted to set up a rapid transformation method using the change of salt concentrations for D. salina. Based on osmotic pressure difference, exogenous genes can be spontaneously transferred into D. salina cells. After that, transformed D. salina cells were subjected to histochemical and molecular analysis. The results showed that the reporter gene, beta-glucuronidase genes were successfully expressed in the positive transformants, and detected in all of transformed cells by PCR analysis. Moreover, different transformation parameters, containing the salt gradient, time, dye dosage and Triton X-100 concentration, were optimized to obtain an optimal transformation result. Taken together, we preliminarily established a rapid transformation method with the features of fast, simple, economic, and high-efficient. This method will provide a strong genetic manipulation tool for the future transformation of D. salina system.

中文翻译：

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杜氏盐藻系统快速转化方法的探索

作为一种新的表达系统，杜氏盐藻（D. salina）在各个领域都具有广阔的前景和应用前景。然而，由于外来基因在盐藻中的低表达和不稳定性，目前其应用受到限制。在遗传操作过程中，转化是盐藻系统中基因表达的关键步骤。尽管当前存在几种转换方法，但是在实际实践中仍然可以发现许多固有的缺陷和局限性。因此，我们试图建立一种利用盐藻盐浓度变化的快速转化方法。根据渗透压差，可以将外源基因自发转移到D.盐藻细胞。之后，对转化的盐藻D. salina细胞进行组织化学和分子分析。结果表明，报告基因，β-葡糖醛酸糖苷酶基因在阳性转化体中成功表达，并通过PCR分析在所有转化细胞中检测到。此外，优化了不同的转化参数，包括盐梯度，时间，染料用量和Triton X-100浓度，以获得最佳的转化结果。综上所述，我们初步建立了一种快速，简单，经济，高效的快速转换方法。该方法将为D. salina系统的未来转化提供强大的遗传操作工具。