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粉棒束孢是一种常见的昆虫病原真菌,被广泛用于生物防治。运用PCR技术与DNA步移技术,从粉棒束孢中克隆出类枯草杆菌蛋白酶的结构基因及其上游序列。结构基因总长1 650 bp,扩出的结构基因DNA与其cDNA比较在距离起始密码子369 bp处有一个内含子,大小为51 bp。通过DNA步移技术得到的上游序列大小为3 189 bp,与cDNA相互重叠的部分为352 bp。分析表明,上游序列中没有明显的TATA-盒和CAAT-盒,但含有GC-box、热激转录因子(HSF)等重要的转录因子结合位点,以及GATA等启动子顺式调控元件。
Conidium is a common entomopathogenic fungus that is widely used in biological control. Using PCR and DNA walking techniques, the structural genes of subtilisin-like protease and their upstream sequences were cloned from the genus Conidia. The total length of the structural gene is 1 650 bp. The amplified structural gene DNA has an intron at a distance of 369 bp from the start codon and has a size of 51 bp. The size of the upstream sequence obtained by DNA walking was 3 189 bp and the overlap with cDNA was 352 bp. The analysis showed that there is no obvious TATA-box and CAAT-box in the upstream sequence, but contains important transcription factor binding sites such as GC-box and heat shock transcription factor (HSF), and cis-regulatory elements of GATA and other promoters.