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以玉米大斑病菌为材料,根据同源序列法分别进行了3′端和5′端的RACE扩增,获得了玉米大斑病菌钙调素基因cDNA全长。利用genomic walking技术获得了CaM基因启动子序列,该序列含有多个与转录调控有关的保守序列(如TATA-box、Sp1、AP-2和TFIID)。Southern杂交结果表明,玉米大斑病菌CaM基因在基因组中以单拷贝形式存在。用CaM特异性抑制剂三氟拉嗪(trifluoperazine)处理玉米大斑病菌,发现其对孢子萌发和附着胞形成的抑制作用与浓度呈正相关,同一浓度下,抑制剂对附着胞形成的抑制作用大于对孢子萌发的抑制。推测CaM基因在玉米大斑病菌的致病过程中起着至关重要的作用。
According to homologous sequence method, RACE amplification of 3 ’end and 5’ end were carried out to obtain the full length cDNA of calmodulin gene from S. turcica. The genomic walking technique was used to obtain the CaM gene promoter sequence, which contains several conserved sequences related to transcriptional regulation (such as TATA-box, Sp1, AP-2 and TFIID). Southern hybridization results showed that the CaM gene of S. turcica was present as a single copy in the genome. The CaM-specific inhibitor trifluoperazine was used to treat S. turcica and its inhibitory effect on the spore germination and attachment formation was positively correlated with its concentration. At the same concentration, the inhibitor inhibited the attachment formation more than Inhibition of spore germination. It is speculated that the CaM gene plays a crucial role in the pathogenesis of S. turcica.