Visualization of nuclei in Aspergillus oryzae with EGFP and analysis of the number of nuclei in each conidium by FACS
Visualization of nuclei in Aspergillus oryzae with EGFP and analysis of the number of nuclei in each conidium by FACS
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DOI：
10.1271/bbb.65.1504
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发表时间：
2001-07-01
影响因子：
1.6
通讯作者：
Kitamoto, K
中科院分区：
文献类型：
--
作者：
Maruyama, J;Nakajima, H;Kitamoto, K
据报道，黑曲霉可形成多核分生孢子。为了分析活细胞中的细胞核，我们开发了A. nidulans histone H2B protein taged by EGFP（H2B：：EGFP）. A. A300和A.在表达H2B：：EGFP的nidulans FGSC 89转化子中，在菌丝和分生孢子的核区域检测到荧光。而A. nidulans nidulans nidulans nidulans中，约有66%的分生孢子有两个，24%的分生孢子有一个，10%的分生孢子有三个或三个以上。好吧将表达H2B：：EGFP的分生孢子进行FACS（荧光激活细胞分选）分析，在A.好吧此外，A.用流式细胞仪成功分离出的单核分生孢子，其核数分布与原始分生孢子的核数分布几乎相同。五个A的分生孢子。对清酒酿造中使用的酿酒酵母菌株的核数进行了评分，结果表明每个分生孢子中存在不同数量的核，有些菌株有少量的单核分生孢子。
Aspergillus oryzae has been reported to form conidia with multinuclei. In order to analyze nuclei in living cells, we developed an expression system of the A. nidulans histone H2B protein tagged by EGFP (H2B::EGFP). In both A. oryzae niaD300 and A. nidulans FGSC89 transformants expressing H2B::EGFP, fluorescence was detected in nuclear regions of hyphae and conidia. While a conidium contained only one fluorescent spot in the A. nidulans transformant, approximately 66% of conidia had two, 24% had one, and 10% had three or more in the A. oryzae transformant. The conidia expressing H2B::EGFP were put through FACS (fluorescence-activated cell sorting) analysis and two sharp peaks, corresponding to one and two nuclei in each conidium, were noted in the A. oryzae transformant. In addition, the A. oryzae uninucleate conidia that were successfully isolated by FACS reproduced conidia with almost the same number distribution of nuclei as that of the original. Conidia of five A. oryzae strains used in sake brewing were scored for the number of nuclei, showing that a varied number of nuclei existed in each conidium and some strains had a small number of uninucleate conidia.