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酿酒酵母单倍体筛选及鉴定方法比较
发表时间：2014-03-15 浏览量：2127 下载量：780
| 全部作者： | 张金金，黄广庆，李智斌，林蒋海，龚映雪，李晶博，肖文娟，刘泽寰 |
| 作者单位： | 暨南大学生命科学技术学院；广东中科天元新能源科技有限公司 |
| 摘 要： | 目的：从工业酿酒酵母AS2.489诱导产孢分离筛选单倍体，比较5种用于鉴定单倍体的方法，以求简易准确地鉴定出可用于后期分子生物学改造的单倍体菌株。方法：分别用生长形态比较法，酵母杂交鉴定法，基因组特异性PCR，DNA含量比较法和胁迫条件耐受性鉴定法对筛选的单倍体进行鉴定，比较这5种方法的差异。结果：生长形态比较法、DNA含量鉴定法和胁迫条件耐受性鉴定法适合初步区分单倍体和二倍体，不适合单倍体配型的鉴定。酵母杂交鉴定法、基因组特异性PCR实验结果稳定准确。筛选得到 3株稳定单倍体MATa配型AS-5和AS-21菌株，MATα配型AS-7菌株。结论：将任意4种方法与酵母杂交鉴定法结合，可筛选得到稳定单倍体菌株。 |
| 关 键 词： | 微生物学；酿酒酵母；单倍体；筛选；鉴定 |
| Title： | Comparison of haploid strain selection of Saccharomyces cerevisiae and identification methods |
| Author： | ZHANG Jinjin, HUANG Guangqing, LI Zhibin, LIN Jianghai, GONG Yingxue, LI Jingbo, XIAO Wenjuan, LIU Zehuan |
| Organization： | College of Life Science and Technology, Jinan University; Guangdong Zhongke Tianyuan New Energy Technology Co., Ltd. |
| Abstract： | Objective: To evaluate the five methods of identification haploid strain for molecular biology research. The haploid strains of AS line from the industrial strain AS2.489 of Saccharomyces cerevisiae have been obtained by sporulation. Methods: The five methods including comparison of the morphological characteristic of diploid and haploid, cross breeding, specific PCR, concentration analysis genomic DNA, andstress tolerance of ethanol and glucose osmotic pressure were used and their difference was comparied. Results: With the regard to the stability and accuracy of haploid strain, the cross breeding and specific PCR were more stable than the other three. others were able to be applied to separate haploid strain from diploid strain. Three stable haplotype strains (two MATa, AS-5 and AS-21, and one MATα AS-7) were separated in this study. Conclusion: One of other methods combined with cross breeding may be suitable for selection of haploid strain from yeast diploid. |
| Key words： | microbiology; Saccharomyces cerevisiae; haploid; selection; identification |
| 发表期数： | 2014年3月第5期 |
| 引用格式： | 张金金，黄广庆，李智斌，等. 酿酒酵母单倍体筛选及鉴定方法比较[J]. 中国科技论文在线精品论文，2014，7（5）：428-434. |
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