Aspergillus oryzae SAICAR合成酶基因的克隆及反义表达载体的构建

Cloning of SAICAR Synthetase Gene of Aspergillus oryzae and Construction of Antisense Expression Vector

目的 构建Aspergillus oryzae新的红色表现型菌株用于天然色素生产。方法 构建Aspergillus oryzaeRIB40 cDNA文库,应用BLAST网络服务对Aspergillus oryzae RIB40 cDNA文库的EST数据进行同源性比较,从EST克隆扩增Aspergillus oryzae类SAICAR合成酶基因(sh1919f片段),反向插入pUSA真核表达载体。结果 完成反义表达载体的构建。Aspergillus oryzae类SAICAR合成酶氨基酸序列与Pichia jadinii、Saccharomyces cerevisiae和Schizosac-charomyces pombe的SAICAR合成酶氨基酸序列具有高度同源性。结论 类SAICAR合成酶基因反义表达载体的构建为其转染入Aspergillus oryzae菌株筛选红色突变菌株奠定了实验基础。

Objective To establish a new red phenotype strain of Aspergillus oryzae for manufacturing natural pigments. Methods Construct the cDNA library of Aspergillus oryzae RIB40 and compare the homolo-gy of EST data in the library using BLAST program. Amplify the SAICAR synthetase gene fragment sh1919f of Aspergillus oryzae and insert into pUSA eukaryotic expression vector at reverse direction. Results An anti-sense expression vector was constructed. The amino acid frequence deduced from the DNA sequence of SAICAR synthetase gene of Aspergillus oryzae was highly homologous to those of Pichia jadinii, Saccharomyces cerevisiae and Schizosaccharomyes pombe. Conclusion The construction of antisense expression vector of SAICAR syn-thelase gene laid a foundation of screening red phenotype strain of Aspergillus oryzae .