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首页> 外文会议>International Conference on Mushroom Biology and Mushroom Products >Transformation of Volvariella volvacea with a Thermal Hysteresis Protein Gene by Particle Bombardment
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Transformation of Volvariella volvacea with a Thermal Hysteresis Protein Gene by Particle Bombardment

机译：通过粒子轰击将Volvariella volvacea与热滞后蛋白基因的转化

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A cDNA encoding a thermal hysteresis protein gene was isolated from the Arctic insect spruce budworm by RT-PCR amplification. It was used to transform Volvariella volvacea through particle bombardment. PCR detection and Southern blotting analysis showed that the thermal hysteresis protein gene was integrated into V. volvacea genomes. A cold stress assay revealed that transgenic V. volvacea exhibited stronger cold tolerance than the host strain. The properties of transgenic V. volvacea were also changed in terms of growth rates and hyphal morphology. Growth rates of most V. volvacea transformants were significantly slower than those of a negative control strain. Furthermore, hyphae of most V. volvacea transformants were thinner than hyphae of the host strain. Experimental results also showed that V. volvacea transformants resistant to hygromycin were mitotically stable. Three-rounds of selection with the first selection on PDSA solid selective medium followed by second and third selections in PDSB liquid selective medium was the best selection procedure for V. volvacea transformant screening.
机译：通过RT-PCR扩增，从北极昆虫云芽虫中分离了编码热滞后蛋白基因的cDNA。它用于通过粒子轰击转化volvariella volvacea。 PCR检测和Southern印迹分析表明，热滞后蛋白基因集成到V.Volvacea基因组中。冷应激测定显示，转基因V.Volvacea表现出比宿主菌株更强的耐寒性。转基因V.Volvacea的性质也在生长率和亚腿形态方面发生变化。大多数V. Volvacea转化体的生长率明显慢于阴性对照菌株。此外，大多数V.Volvacea转化体的菌丝比宿主菌株的菌丝稀释。实验结果还表明，抗潮霉素的Volvacea转化体是显着的。在PDSA固体选择性培养基上的第一个选择三轮选择，然后在PDSB液体选择性培养基中进行第二和第三选择是V.Volvacea转化体筛选的最佳选择过程。

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