摘要
枯草芽孢杆菌ZA1具有良好的抑菌和促生作用。解旋酶已被证实为核酸代谢关键蛋白复合体,且解旋酶超家族成员通过特异性切割DNA/RNA分子中的磷酸二酯键,在遗传信息传递过程中执行关键催化功能。为明确解旋酶基因ypvA和yjcD在枯草芽孢杆菌ZA1抑菌过程中的潜在作用,以枯草芽孢杆菌ZA1为材料,克隆ypvA和yjcD基因,利用生物信息学分析它们的序列特征,通过亚细胞定位检测2个基因的位置,对2个基因及其编码蛋白的结构与功能特性进行初步解析。结果表明,克隆获得的ypvA和yjcD基因编码区全长分别为1791,1767 bp,分别编码596,588个氨基酸,其编码蛋白的预测相对分子质量分别为68.80401,68.27508 ku,理论等电点分别为4.81,8.25,且亲水性较弱。ypvA和yjcD的蛋白空间结构主要由α-螺旋、β-折叠和大量无规则延伸链组成的紧密结构。亚细胞定位于细胞质和细胞核中,不存在跨膜区及信号肽,为非分泌蛋白。分析保守结构域得到ypvA属于Rad3相关DNA解旋酶DinG家族,基因yjcD属于UvrD超家族I DNA或RNA解旋酶,通过进一步的蛋白质系统进化树分析明确将这2个蛋白鉴定为ATP依赖性解旋酶。因此,通过克隆枯草芽孢杆菌ZA1中的ypvA和yjcD基因并分析序列为非分泌性ATP依赖性解旋酶。
Bacillus subtilis ZA1 exhibits notable antibacterial and growth-promoting activities.Helicases have been established as crucial protein complexes in nucleic acid metabolism,with members of the helicase superfamily performing key catalytic functions during genetic information transfer by specifically cleaving phosphodiester bonds in DNA/RNA molecules.To elucidate the potential roles of the helicase genes ypvA and yjcD in the antibacterial process of B.subtilis ZA1,we used B.subtilis ZA1 as the experimental material,cloned the ypvA and yjcD genes,analyzed their sequence characteristics through bioinformatics,and determined their subcellular localization.A preliminary investigation was conducted on the structural and functional properties of these two genes and their encoded proteins.The results showed that the cloned coding sequences(CDS)of ypvA and yjcD were 1791,1767 bp in length,encoding 596,588 amino acids,respectively.The predicted relative molecular masses of the encoded proteins were 68.80401,68.27508 ku,with theoretical isoelectric points(pI)of 4.81,8.25,respectively,and both exhibited weak hydrophilicity.The spatial structures of the ypvA and yjcD proteins consisted of a compact architecture primarily composed ofα-helices,β-sheets,and extensive random coils.Subcellular localization analysis indicated that both proteins were localized in the cytoplasm and nucleus,lacking transmembrane domains and signal peptides,thus classifying them as non-secretory proteins.Analysis of conserved domains revealed that ypvA belonged to the DinG family of Rad3-related DNA helicases,while yjcD belongs to the UvrD superfamily I DNA/RNA helicases.Further phylogenetic analysis confirmed that both proteins are ATP-dependent helicases.In summary,cloning and sequence analysis of the ypvA and yjcD genes from B.subtilis ZA1 identified them as non-secretory ATP-dependent helicases.
作者
任姝锦
蔡锋锋
马婷
王婷
施玉安
王一丹
杨成德
REN Shujin;CAI Fengfeng;MA Ting;WANG Ting;SHI Yu′an;WANG Yidan;YANG Chengde(Laboratory of Biocontrol Engineering of Crop Pests and Diseases in Gansu Province,College of Plant Protection,Gansu Agricultural University,Lanzhou 733070,China)
出处
《华北农学报》
北大核心
2025年第S1期285-291,共7页
Acta Agriculturae Boreali-Sinica
基金
甘肃省重点研究项目(24ZDNA009-2)。
关键词
枯草芽孢杆菌
解旋酶基因
克隆
生物信息分析
亚细胞定位
Bacillus subtilis
Helicase gene
Cloning
Bioinformation analysis
Subcellular localization
