抗冷冻蛋白基因遗传转化草菇的研究
Transformation of Volvariella volvacea with a thermal hysteresis protein gene by particle bombardment
郭丽琼 林俊芳 熊盛 陈守才
摘 要：采用RT-PCR技术从瑞典的北极云杉卷叶蛾幼虫中扩增出抗冷冻蛋白基因,利用基因枪法遗传转化草菇.PCR检测和Southem杂交结果证明,抗冷冻蛋白基因已整合进草菇基因组.低温胁迫试验结果表明,转基因草菇具有较强的耐低温能力.转基因草菇生物学特性观测结果显示,大多数草菇转化子的生长速率明显地慢于对照的宿主菌株,多数转化子的菌丝也明显地比宿主菌丝细弱.转化子筛选结果表明,采用三轮的转化子筛选程序,即第一轮在固体培养基上筛选、第二轮和第三轮在液体培养基中筛选,有利于获得真实转化子和淘汰假转化子.转基因草菇一代低温胁迫结果证明,转基因草菇后代仍然具有较强的低温耐受能力,这说明转基因草菇的耐低温性能在世代之间是稳定的.
关键词：草菇,抗冷冻蛋白基因,转化,基因枪法
分类号：S66.13,Q939.96 文献标识码：A
文章编号：0001-6209(2005)01-0039-05
基金项目：国家自然科学基金(39960050,30060054,30371000);广东省自然科学基金(032239);华南农业大学校长基金(5100-K03003)
作者简介：郭丽琼(1963-),女,福州市人,副研究员,博士,主要从事大型真菌分子生物学和生物技术研究.E-mail:guolq@scau.edu.cn.林俊芳,通讯作者.Tel:86-20-38902762;Fax:86-20-85280270;E-mail:linjf@scau.edu.cn
作者单位：郭丽琼（华南农业大学食品学院,生物工程系,广州,510640;中国热带农业科学院,热带作物生物技术国家重点实验室,海口,571101）
林俊芳（华南农业大学食品学院,生物工程系,广州,510640;中国热带农业科学院,热带作物生物技术国家重点实验室,海口,571101）
熊盛（中国热带农业科学院,热带作物生物技术国家重点实验室,海口,571101）
陈守才（中国热带农业科学院,热带作物生物技术国家重点实验室,海口,571101）
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微生物学报
ACTA MICROBIOLOGICA SINICA
2005 Vol.45 No.1 P.39-43
