抗冷凍蛋白基因遺傳轉化草菇的研究
Transformation of Volvariella volvacea with a thermal hysteresis protein gene by particle bombardment
郭麗瓊 林俊芳 熊盛 陳守才
摘 要：采用RT-PCR技術從瑞典的北極云杉卷葉蛾幼蟲中擴增出抗冷凍蛋白基因,利用基因槍法遺傳轉化草菇.PCR檢測和Southem雜交結果證明,抗冷凍蛋白基因已整合進草菇基因組.低溫脅迫試驗結果表明,轉基因草菇具有較強的耐低溫能力.轉基因草菇生物學特性觀測結果顯示,大多數草菇轉化子的生長速率明顯地慢于對照的宿主菌株,多數轉化子的菌絲也明顯地比宿主菌絲細弱.轉化子篩選結果表明,采用三輪的轉化子篩選程序,即第一輪在固體培養基上篩選、第二輪和第三輪在液體培養基中篩選,有利于獲得真實轉化子和淘汰假轉化子.轉基因草菇一代低溫脅迫結果證明,轉基因草菇后代仍然具有較強的低溫耐受能力,這說明轉基因草菇的耐低溫性能在世代之間是穩定的.
關鍵詞：草菇,抗冷凍蛋白基因,轉化,基因槍法
分類號：S66.13,Q939.96 文獻標識碼：A
文章編號：0001-6209(2005)01-0039-05
基金項目：國家自然科學基金(39960050,30060054,30371000);廣東省自然科學基金(032239);華南農業大學校長基金(5100-K03003)
作者簡介：郭麗瓊(1963-),女,福州市人,副研究員,博士,主要從事大型真菌分子生物學和生物技術研究.E-mail:guolq@scau.edu.cn.林俊芳,通訊作者.Tel:86-20-38902762;Fax:86-20-85280270;E-mail:linjf@scau.edu.cn
作者單位：郭麗瓊（華南農業大學食品學院,生物工程系,廣州,510640;中國熱帶農業科學院,熱帶作物生物技術國家重點實驗室,?？?571101）
林俊芳（華南農業大學食品學院,生物工程系,廣州,510640;中國熱帶農業科學院,熱帶作物生物技術國家重點實驗室,?？?571101）
熊盛（中國熱帶農業科學院,熱帶作物生物技術國家重點實驗室,?？?571101）
陳守才（中國熱帶農業科學院,熱帶作物生物技術國家重點實驗室,?？?571101）
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微生物學報
ACTA MICROBIOLOGICA SINICA
2005 Vol.45 No.1 P.39-43
