大肠埃希菌中细胞分裂抑制子的潜在调控基因筛选

Screening of potential regulatory genes of Sdi A in Escherichia coli

目的筛选大肠埃希菌中细胞分裂抑制子(Sdi A)的潜在调控基因。方法将大肠埃希菌E.coli SM10λpir野生株、E.coli SM10λpir Sdi A敲除株、E.coli SM10λpir Sdi A过表达回复株分别加入N-酰基-L-高丝氨酸内酯(AHLs)信号分子处理,记为AHLs处理组;对应三种菌株分别加入等量DMSO处理,记为非AHLs处理组。两组菌株基因转录组测序后,通过R语言edgeR软件包筛选差异表达基因。选取两组内E.coli SM10λpir Sdi A敲除株与E.coli SM10λpir野生株之间、E.coli SM10λpir Sdi A过表达回复株与E.coli SM10λpir Sdi A敲除株之间同时出现差异性表达的基因,即为有/无AHLs信号分子时Sdi A的潜在调控基因,并进行组间比对,观察有/无AHLs信号分子时Sdi A潜在调控基因的变化。结果非AHLs处理组中,E.coli SM10λpir Sdi A敲除株与E.coli SM10λpir野生株之间、E.coli SM10λpir Sdi A过表达回复株与E.coli SM10λpir Sdi A敲除株之间同时出现差异性表达的基因为nar H、nar J、nar I、yee E、yeeR、paa E、flu。AHLs处理组中,E.coli SM10λpir Sdi A敲除株与E.coli SM10λpir野生株之间、E.coli SM10λpir Sdi A过表达回复株与E.coli SM10λpir Sdi A敲除株之间同时出现差异性表达的基因为hcp、ytf E、hcr、hmp、nar H、nar J、nar I、yee E、yeeR、flu、ydi V、hdh A。nar H、nar J、nar I、yee E、yeeR、flu基因在两组中均为Sdi A的潜在调控基因,hcp、ytf E、hcr、hmp、ydi V、hdh A基因仅在AHLs信号分子存在时为Sdi A的潜在调控基因。结论大肠埃希菌中,Sdi A的潜在调控基因有nar H、nar J、nar I、yee E、yeeR、flu、paa E、hcp、ytf E、hcr、hmp、ydi V、hdh A,其中对hcp、ytf E、hcr、hmp、ydi V、hdh A基因的调控依赖于AHLs信号分子。

Objective To screen out the potential regulatory genes of suppressor of cell division(Sdi A)in Escherichia coli.Methods E.coli SM10λpir wild strain,E.coli SM10λpir Sdi A mutant strain,and E.coli SM10λpir Sdi A overexpression strain were treated with N-acyl-L-homoserine lactones(AHLs)signal molecules and recorded as the AHLs treatment group.On the other hand,the three strains were treated with equal amount of DMSO,which were recorded as the non-AHLs treatment group.After all the strains were sequenced,the differentially expressed genes were screened out by edgeR software.We selected the common differentially expressed genes between E.coli SM10λpir Sdi A mutant strain and E.coli SM10λpir wild strain and between the E.coli SM10λpir Sdi A overexpression strain and E.coli SM10λpir Sdi A mutant strain as the potential regulatory genes of Sdi A with or without AHLs signaling molecules,and observed the changes in the potential regulatory genes of Sdi A with or without AHLs signaling molecules.Results In the non-AHLs treatment group,the common differentially expressed genes between E.coli SM10λpir Sdi A mutant strain and E.coli SM10λpir wild strain and between the E.coli SM10λpir Sdi A overexpression strain and E.coli SM10λpir Sdi A mutant strain were nar H,nar J,nar I,yee E,yeeR,paa E and flu.In the AHLs treatment group,the common differentially expressed genes between E.coli SM10λpir Sdi A mutant strain and E.coli SM10λpir wild strain,and between the E.coli SM10λpir Sdi A overexpression strain and E.coli SM10λpir Sdi A mutant strain were hcp,ytf E,hcr,hmp,nar H,nar J,nar I,yee E,yeeR,flu,ydi V and hdh A.According to the comparison,nar H,nar J,nar I,yee E,yeeR and flu genes were the potential regulatory genes of Sdi A,and meanwhile,hcp,ytf E,hcr,hmp,ydi V,hdh A genes were the potential regulatory genes of Sdi A in the presence of AHLs signaling molecules.Conclusion In Escherichia coli,the potential regulatory genes of Sdi A are nar H,nar J,nar I,yee E,yeeR,flu,paa E,hcp,ytf E,hcr,hmp,ydi V,and hdh A,and the regulation on hcp,ytf E,hcr,hmp,ydi V and hdh A gene still mainly depends on AHLs signaling molecules.