Investigating the effectiveness of DNA microarray analysis for identifying the genes involved in l-lactate production by Saccharomyces cerevisiae
Investigating the effectiveness of DNA microarray analysis for identifying the genes involved in l-lactate production by Saccharomyces cerevisiae
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DOI：
10.1007/s00253-009-2209-z
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发表时间：
2009-09
影响因子：
5
通讯作者：
T. Hirasawa;A. Ookubo;Katsunori Yoshikawa;K. Nagahisa;C. Furusawa;H. Sawai;H. Shimizu
中科院分区：
文献类型：
--
作者：
T. Hirasawa;A. Ookubo;Katsunori Yoshikawa;K. Nagahisa;C. Furusawa;H. Sawai;H. Shimizu
为了确定通过DNA微阵列分析获得的转录组数据是否可用于鉴定参与靶代谢产物产生的基因，我们试图通过产乳酸重组酵母菌株鉴定参与l-乳酸产生的基因。我们获得了这些菌株的DNA微阵列数据。将携带乳酸菌、牛和人L-乳酸脱氢酶（LDH）基因的质粒导入PDC 1-破坏的S。携带人和牛LDH基因的菌株的产L-乳酸能力高于携带乳酸菌LDH基因的菌株。DNA微阵列分析显示，388个基因的表达显着改变的菌株与人和牛LDH基因。其中，289个基因的人LDH-携带的缺失菌株的乳酸生产力与标准和56个随机选择的含有相同LDH基因的缺失菌株的乳酸生产力进行了比较，以验证DNA微阵列分析用于鉴定重组菌株中负责L-乳酸生产的基因的有效性。与标准和随机选择的缺失菌株相比，只有基于DNA微阵列数据选择的基因的缺失菌株显示出显著改变的l-乳酸盐生产。我们的研究结果表明，可以成功地确定相关基因的tol-lactate生产通过选择基因的表达显着改变的DNA芯片分析，和DNA芯片分析的有效性，确定负责l-乳酸生产的基因进行了讨论。
In order to determine whether transcriptome data obtained by DNA microarray analysis could be used to identify the genes involved in target metabolite production, we tried to identify the genes involved inl-lactate production byl-lactate-producing recombinantSaccharomyces cerevisiaestrains. We obtained DNA microarray data for these strains. Plasmids carrying lactic acid bacteria, bovine, and humanl-lactate dehydrogenase (LDH) genes were introduced intoPDC1-disruptedS. cerevisiaestrains.l-Lactate productivity of the strains harboring the human and bovine LDH genes was higher than that of the strains harboring lactic acid bacteria LDH genes. DNA microarray analysis revealed that the expression of 388 genes was significantly altered in the strains with the human and bovine LDH genes. Of these, thel-lactate productivity of human LDH-harboring deletion strains of 289 genes was compared with that of the standard and 56 randomly selected deletion strains containing the same LDH gene to validate the effectiveness of DNA microarray analysis for identifying the genes responsible forl-lactate production in the recombinant strains. Only deletion strains of the genes selected on the basis of the DNA microarray data showed significantly alteredl-lactate production as compared to the standard and the randomly selected deletion strains. Our results indicated that the genes related tol-lactate production could be successfully identified by selecting the genes that exhibited significantly altered expression on DNA microarray analysis, and the effectiveness of DNA microarray analysis for identifying the genes responsible forl-lactate production was discussed.