应用电击法获得转MT基因平菇_英文_.pdfVIP

A cta Botanica Sinica 2002, 44 ( 12) : 1445- 1449 Transformation of Metallothionein Gene into Mushroom Protoplasts by Application of Electroporation * ZHANG Jing, N IE Si_Wei, SHAN Long, RU Bing_Gen (National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Lif e Sciences , Peking University, Beijing 10087 1, China Abstract: Metallothionein gene ( MT has been transferred into mushroom protoplasts by electroporat ion. It is a low molecular weight , cysteine_rich and metal_binding protein. MT can bind metals. Its synthesis is in duced by Zn ion . Thus the expression of MT gene in mushroom can improve the accumulat ion of Zn in this fun gus. This transgenic mushroom, consumed as a kind of vegetable, can supply the necessary Zn to people who 6 are short of the element. When protoplasts were prepared , the concentration ( C of protoplasts is 6. 745 10 / mL. After protoplast electroporat ion , the transformation rate of protoplasts is 0. 01% . Polymerase chain reac tion (PCR analysis showed that the gene had been integrated into the mushroom chromosome. SDS_PAGE, Western blot analysis indicated that the MT gene had been expressed in the transgenic mushroom. The express ing level, detected by ELISA, is 0. 6% - 0. 8% . T ested for metal resistance, the wild_type mushroom growth was inhibited on the medium containing 1. 0 - 1. 2 mmol/ L ZnSO4. While the transgenic mushroom was inhib ited on the medium