College of Forestry, Northeast Forestry University, Hexing Road 26, Xiangfang District, Harbin, 150040, Heilongjiang, China.
College of Life Science, Northeast Forestry University, Hexing Road 26, Xiangfang District, Harbin, 150040, Heilongjiang, China.
Protein Expr Purif. 2020 Jun;170:105592. doi: 10.1016/j.pep.2020.105592. Epub 2020 Feb 5.
Acetyl-CoA C-acetyltransferase synthase gene (AACT) cDNA, DNA and promoter were cloned from Sanghuangporus baumii. The gene ORF (1260 bp) encoded 419 amino acids. The AACT DNA includes five exons (1-84 bp, 140-513 bp, 570-1027 bp, 1090-1282 bp, 1344-1494 bp) and four introns (85-139 bp, 514-569 bp, 1028-1089 bp, 1283-1343 bp). The molecular weight of AACT protein is 43.40 kDa, it is hydrophilic with a theoretical isoelectric point of 8.96. Furthermore, The region of the transcription start site is 1997-2047 bp of AACT promoter, and it contained promoter elements (TATA Boxs, CAAT Boxs, CAAT-box, ABRE, G-Boxs, Sp1, MSA-like, LTR). AACT recombinant protein (43.40 KDa + Tag protein 22.68 KDa) was subjected in SDS-PAGE. AACT the transcription levels of in different development stages were investigated. The expression of AACT in primordia (2.4-fold) and 15 d mycelia (2.3- fold) were significantly higher than 9 d mycelia (contral). The expression level of the AACT downstream genes and triterpenoids content were determined at different developmental stages. Triterpenoid content reached its peak on day 15(7.21 mg/g).
从桑黄中克隆得到乙酰辅酶 A C-乙酰转移酶合酶基因（AACT）的 cDNA、DNA 和启动子。该基因的 ORF（1260 bp）编码 419 个氨基酸。AACT DNA 包含五个外显子（1-84 bp、140-513 bp、570-1027 bp、1090-1282 bp、1344-1494 bp）和四个内含子（85-139 bp、514-569 bp、1028-1089 bp、1283-1343 bp）。AACT 蛋白的分子量为 43.40 kDa，具有亲水性，理论等电点为 8.96。此外，AACT 启动子转录起始位点区域为 1997-2047 bp，包含启动子元件（TATA 盒、CAAT 盒、CAAT 框、ABRE、G-Boxs、Sp1、MSA-like、LTR）。AACT 重组蛋白（43.40 kDa+Tag 蛋白 22.68 kDa）在 SDS-PAGE 中进行了分析。研究了不同发育阶段 AACT 的转录水平。原基（2.4 倍）和 15 d 菌丝（2.3-2.3 倍）中的 AACT 表达明显高于 9 d 菌丝（对照）。在不同发育阶段测定了 AACT 下游基因和三萜含量的表达水平。三萜含量在第 15 天达到峰值（7.21 mg/g）。
