| 酿酒酵母染色体DNA样品制备和CHEF分析 |
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| 引用本文： | 边银丙 翁曼丽 罗信昌 金德敏 周启 王斌. 酿酒酵母染色体DNA样品制备和CHEF分析[J]. 菌物学报, 1999, 18(3) |
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| 作者姓名： | 边银丙 翁曼丽 罗信昌 金德敏 周启 王斌 |
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| 作者单位： | 1.华中农业大学应用真菌研究所 武汉 430070；2.华中农业大学生命科技学院 武汉 430070；3.中国科学院遗传研究所 北京 100101 |
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| 基金项目： | 中国科学院遗传所植物生物技术开放实验室资助 |
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| 摘 要： | 等高锁状均匀电场（contolllsclampedhomogeneouselectncfield，CHED电泳技术与分子杂交、酵母人工染色体（yeastartificialchromosome，YAC）等分子生物学技术的有机结合，推动了真菌电泳核型、染色体作图和染色体DNA长度多型性（chromosomalDNAlellgthpolymorphism，CLP...
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| 关 键 词： | 酿酒酵母 原生质体 染色体DNA样品 CHEF分析 |
PREPARATION AND CHEF ANALYSIS OF YEAST CHROMOSOMAL DNA SAMPLE |
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| BIAN Yin--Bing, WENG Man--Li, LUO Xin--ChangJIN De--MIN, ZHOU Qi, WANG Bin. PREPARATION AND CHEF ANALYSIS OF YEAST CHROMOSOMAL DNA SAMPLE[J]. Mycosystema, 1999, 18(3) |
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| Authors: | BIAN Yin--Bing WENG Man--Li LUO Xin--ChangJIN De--MIN ZHOU Qi WANG Bin |
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| Affiliation: | BIAN Yin--Bing; WENG Man--Li; LUO Xin--ChangJIN De--MIN; ZHOU Qi; WANG Bin |
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| Abstract: | 1.5% Lywallzyme (product of Guangdong Microbiology Institute, China) was used to remove the cell walls of Saccharomyces cerevisae in the preparation of protoplasts. The protoplasts resuspented in ST buffer at the concentration of 4 × 108 / ml were mixed with an equal volume of 1.0% low-melting point agarose in ST buffer at 50℃. In order to prepare yeast chromosomal DNA samples, the chilled agarose Plugs were incubated in NDS buffer containing ling/ml proteinase K for 24h. This procedure of chromosomal DNA sample prepeon,which is different from that adopted by former researchers, have laid a fundation for the preparation of megabase-size DNA marker and electrophorehc karyotype analysis of other fungi. It was revealed by CHEF electrophoresis that chromosomal DNA length polymorphism exists between tested stain X8 and control stain ThH755. |
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| Keywords: | Saccharomyces cerevisae Protoplast Chromosomal DNA sample CHEF electrophoresis |
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