球孢白僵菌tps2基因的克隆和生物信息学分析

Cloning and Bioinformatics Analysis of Tps2 Gene from Beauveria bassiana

运用SMART RACE RT-PCR技术与DNA步移技术,首次从球孢白僵菌中克隆出完整的海藻糖-6-磷酸磷酸酯酶TPS2的基因编码区序列及上游序列。该基因cDNA全长3 219 bp,其中开放阅读框(ORF)2 619 bp,编码872个氨基酸。成熟蛋白理论分子量为97.8 kD,理论等电点为6.32。编码框结构基因的全长为2 821 bp,有两个长度分别为140 bp和62 bp的内含子。分析表明,上游序列中含有TATA-box、CAAT-box和GC-box,并且也存在GATA元件等启动子顺式调控元件。本文结果将为进一步研究海藻糖在虫生真菌中的生理合成以及抗逆调控机制奠定坚实的基础。

The full-length cDNA of Bbtps2 gene was cloned from Beauveria bassiana using SMART RACE RT-PCR. And its upstream sequence was amplified using genome walking. Analysis of the cloned complete cDNA with a whole length of 3219 bp showed that it encompassed a open reading frame （ORF） with 2619 bp encoding 872 amino acid. The mature protein had a molecular mass of 97.8 kD with a calculated pI of 6.32. The structure gene for Bbtps2 ORF, with a whole sequence of 2821 bp, contains two introns （140 bp and 61 bp, respectively）. The analyses indicate that the upstream sequence contained several regulatory elements, such as TATA-box、CAAT-box、GC-box and GATA. The finding of this study will establish the foundation for further investigating physiological synthesis and anti-adversity regulation of trehalose in entomopathogenic fungi.