| 杜氏盐藻新型遗传转化方法的建立(英文) |
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| 引用本文： | 柴晓杰,靳非,丛玉婷,岳金荣,刘艺琼.杜氏盐藻新型遗传转化方法的建立(英文)[J].农业科学与技术,2017,18(8). |
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| 作者姓名： | 柴晓杰 靳非 丛玉婷 岳金荣 刘艺琼 |
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| 作者单位： | 大连海洋大学/辽宁省水生生物学重点实验室,辽宁大连,116023 |
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| 基金项目： | 国家自然科学基金( 31472260).Supported by National Natural Science Foundation of China |
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| 摘 要： | 本研究采用LiAc/PEG介导法首次成功地将外源基因导入盐藻细胞,组织化学染色发现转基因盐藻为蓝色,表明外源的基因在盐藻细胞中得到了成功表达。同时还进行了生长状态、转化温度、质粒浓度等因素对转化率影响的研究,优化了转化条件。结果表明,盐藻处于对数生长早期、质粒DNA浓度为600μg/ml、转化温度29℃是最理想的转化条件,其转化率为74.8‰。PCR扩增和PCR-Southern杂交结果证明,目的基因已整合到基因组中,且能够遗传。
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| 关 键 词： | 杜氏盐藻 LiAc/PEG介导法 GUS基因 遗传转化 |
Establishment of a New Method for Genetic Transformation of Dunaliella salina |
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| Xiaojie CHAI,Fei JIN,Yuting CONG,Jinrong YUE,Yiqiong LIU.Establishment of a New Method for Genetic Transformation of Dunaliella salina[J].Agricultural Science ＆ Technology,2017,18(8). |
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| Authors: | Xiaojie CHAI Fei JIN Yuting CONG Jinrong YUE Yiqiong LIU |
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| Abstract: | The exogenous gene was integrated into Dunaliella salina successfully by using LiAc/PEG mediating method for the first time. According to the results of histochemical staining, transgenic D. salina was blue, showing that the exogenous GUS gene was successfully expressed in the cells of D. salina. Meanwhile, the effects of growth state of D. salina, plasmid concentration and temperature on its transformation efficiency were studied, and the transformation conditions were optimized. The results show that the optimum conditions for the genetic transformation of D. salina are shown as follows: D. salina was in the early logarithmic phase; plasmid DNA concentration was 600 μg/ml; temperature was 29 ℃, and transformation efficiency was up to 74.8‰ under the best conditions. According to the results of PCR amplification and PCR-Southern hybridization, the target gene had been integrated into genome and was hereditary. |
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| Keywords: | Dunaliela salina LiAc/PEG mediating method GUS gene Genetic transformation |
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