CAJ | 학술논문

本研究运用RACE技术，从球孢白僵菌中克隆出完整的DNA甲基转移酶基因Dim-2的编码区序列。该基因cDNA全长4021 bp，5′端非翻译区283 bp，3′端非翻译区303 bp，开放阅读框（ORF）3429 bp，编码1142个氨基酸。蛋白理论分子量为128 kD，理论等电点为6.13。结构域分析显示，该基因编码蛋白含有1个BAH结构域和合成5-甲基胞嘧啶的活性区域。Real-time PCR与HPLC检测表明，球孢白僵菌DNA甲基化程度会随着其生长发育的变化而不断改变。结果显示，球孢白僵菌Dim-2基因的转录表达量与基因组DNA的5-甲基胞嘧啶百分含量在菌体培养初始产孢阶段（即培养第7 d），均出现了1个低谷。这可能意味着球孢白僵菌Dim-2基因所引起的DNA甲基化与其生长发育之间具有内在的联系。本研究将为进一步探索DNA甲基化在虫生真菌生长发育中具体功能机制奠定基础。
본연구운용RACE기술，종구포백강균중극륭출완정적DNA갑기전이매기인Dim-2적편마구서렬。해기인cDNA전장4021 bp，5′단비번역구283 bp，3′단비번역구303 bp，개방열독광（ORF）3429 bp，편마1142개안기산。단백이론분자량위128 kD，이론등전점위6.13。결구역분석현시，해기인편마단백함유1개BAH결구역화합성5-갑기포밀정적활성구역。Real-time PCR여HPLC검측표명，구포백강균DNA갑기화정도회수착기생장발육적변화이불단개변。결과현시，구포백강균Dim-2기인적전록표체량여기인조DNA적5-갑기포밀정백분함량재균체배양초시산포계단（즉배양제7 d），균출현료1개저곡。저가능의미착구포백강균Dim-2기인소인기적DNA갑기화여기생장발육지간구유내재적련계。본연구장위진일보탐색DNA갑기화재충생진균생장발육중구체공능궤제전정기출。
The full-length cDNA of Dim-2 gene was cloned from Beauveria bassiana using a RACE technique. The cDNA of Dim-2 had a lenght of 4021 bp, including an open reading frame (ORF) with 3429 bp encoding 1142 amino acids. The protein had a molecular mass of 128 kD with a calculated pI of 6.13. The Realtime-PCR and HPLC analysis indicated that the degree of DNA methylation varied along with fungal growth development. The results showed that both the expression level of Dim-2 gene and the degree of DNA methylation reached the minimum levels at initial stage of sporulation (the 7th day). It was inferred that there was an internal relationship between DNA methylation caused by Dim-2 gene and growth development in B. bassiana. The study will benefit future functional studies of DNA methylation in developmental control of entomopathogenic fungi.