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在前期突变体库研究中发现,ybf B缺失的枯草芽胞杆菌Bacillus subtilis BSD-2丧失了其抑真菌活性。本研究通过敲除并恢复BSD-2中ybf B基因,进一步确认其缺失对菌株生物活性的影响,并利用生物信息学手段对其基因序列和编码蛋白质序列进行了预测分析。本研究成功敲除BSD-2菌株的ybf B基因,构建突变株B-Y-k,并成功构建其回复突变株B-Y,平板对峙实验结果显示突变其ybf B基因该菌株失去其抗真菌活性,回复突变株抗真菌活性恢复,表明该基因与其抗真菌相关。生物信息学分析显示该基因全长1 251 bp,编码416个氨基酸,具有12个跨膜螺旋区,为疏水性蛋白,推测该蛋白为疏水性跨膜转运蛋白,可能参与活性物质的转运。
In a previous study of mutant pools, it was found that the ybf B-deficient Bacillus subtilis BSD-2 lost its fungistatic activity. In this study, we knocked out and recovered the ybf B gene in BSD-2 to further confirm the effect of its deletion on the biological activity of the strain, and predicted the gene sequence and the encoded protein sequence by bioinformatics methods. In this study, the ybf B gene of BSD-2 strain was successfully knocked out and the mutant BYk was successfully constructed. The BY strain was successfully constructed. The plate confrontation test results showed that the mutant strain ybf B lost its antifungal activity, Fungal activity recovered, indicating that the gene is associated with its antifungal activity. Bioinformatics analysis showed that the gene was 1 251 bp in length encoding a polypeptide of 416 amino acids with 12 transmembrane helices. It was a hydrophobic protein, suggesting that it is a hydrophobic transmembrane transporter and may be involved in the transport of active substances.