摘要
克隆及鉴定了1株抗肿瘤聚酮去乙酰基真菌环氧二烯(DAM)高产菌株拟茎点霉G818的全局性调控因子veA基因.根据已发现的VeA同源蛋白序列,选取N端保守区域设计简并引物,利用常规PCR及LA-PCR技术获得veA基因序列,然后结合Softberry软件和NCBI数据库分析确定出veA基因的ORF.克隆得到的veA基因,存在1个大小为64 bp的内含子,能够编码381个氨基酸.veA基因的成功克隆为后续RNA干扰菌株的构建及VeA在DAM的生物合成过程中所起作用的探究奠定基础.
The aim of the present paper is to clone a global regulator yeA in a anti-tumor polyketide deacetylmycoepoxydiene （DAM） high-producing strain, Phomopsis sp. GS18. Accord- ing to the known VeA homologous protein sequences, the veA fragment was successfully amplified by PCR using degenerate primers designed by Codehop programme. Then the whole yeA gene sequences were obtained by the gene-specific primer using the LA-PCR technology and confirmed the ORF by means of softberry software and NCBI database. Sequence analysis showed that the yeA ORF has a intron with length of 64 bp and can encode 381 amino acids. This result can be a foundation for the constructing of RNAi strain and the functional analysis of yeA in the biosynthesis of DAM.
出处
《福建师范大学学报（自然科学版）》
CAS
CSCD
北大核心
2014年第2期70-75,共6页
Journal of Fujian Normal University：Natural Science Edition
基金
国家自然科学基金资助项目(31070053)
