菌落PCR快速扩增工业酿酒酵母基因组DNA片段.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 20 10 9 PCR DNA 1, 2, 3 1 1 1 1 1 阳辛凤 高秋芳 李锡敏 郭安平 孔华 贺立卡 1 2 ( , 571101; , 3 570228; , 571101) : 针对工业酿酒酵母细胞破壁难和 取基因酵母组 DNA 费时长( 2- 3 h/样品) 的问题, 研究了 SDS微珠涡旋破 壁的方法以破壁液上清为模板进行菌落PCR扩增酿酒酵母南阳 K基因组中铜抗性蛋白基因( cup1) 片段和 rDNA 片段结 果表明, 在不需要 取酵母基因组前 下, 利用该方法能有效地扩增酵母基因组 DNA 片段, 同时该方法也适用于对转基因酿 酒酵母进行菌落 PCR从而实现对转化子的准确和快速地鉴定筛选, 是一种成本低和易于操作的适于处理大量样品的方法 : 酿酒酵母 菌落 PCR 破壁 Rapid Amplification of Genome DNA Fragment of Industrial Sa ccharomy ces cerevisiae by Colony PCR Y an X infen 1, 2, 3 Gao Q iufan 1 L iXmi in1 Guo Anpin 1 Kon H ua1 H e Lika1 1 ( Institute of Trop icalB ioscience and B iotechnology, CATAS,Haikou 571101; 2 3 AgriculturalCollege,Hainan University,Haikou 570228; Analysis and TestCenter of CATAS,Haikou 571101) A bstrac:t For the d ifficulty of disruptin the cell of industrialSaccharo yces cerevisiae and tmi econsum in ( 2- 3 h / sam ple) to extract yeast enom e, w e studied SDSm in i lass beads vortexm ethod to disrupt the cell of industrialSaccharo yces cerevisiae. Superna tant of cell lysates w as used as temp let for colony PCR to amp lify copperresistant ene CUP 1 fra m ent and ribosom al DNA ( rDNA) fra m en t of Saccharo yces cerevisiae. The resu lts show ed that yeast enome DNA fra m ents could be am plified effectivelyw ithou t extrac tion of yeast enom e. Them ethod wh ich w as proved easy and cheap as performed also cou ld be used for colony PCR to screen the trans form an ts of enetically mod ified yeast accurately and rap idly. K ey w