沙门菌中GcvB sRNA可调控aphA mRNA

GcvB sRNA Regulate aphA mRNA in Salmonella

aphA编码B类酸性磷酸酶,本研究旨在将沙门菌中aphA以lacZ进行替换,获得△aphA::lacZ,从而了解GcvB sRNA对aphA mRNA的调控作用。通过使用λred同源重组、FLP/FRT重组系统、电转导等技术构建相应缺失菌株,并进行β-半乳糖核苷酸酶活实验与qPCR鉴定相应表达量变化。在qPCR实验中,相较于缺失wt、GcvB及其R1、R2、R3后,△aphA::lacZ的aphA相对转录量下降;而在β-半乳糖核苷酸酶活实验中,相较于△aphA::lacZ,缺失GcvB后变化并不明显,而缺失R1、R2、R3后,β-半乳糖苷酶酶活均有上升,分别为244.47%、230.67%、305.06%。以上结果表明,GcvB sRNA对aphA mRNA具有一定的调控作用。

aphA gene can encode class B acid phosphatase/phosphotransferase.The aim of this study was to obtain ΔaphA::lacZ by deleting aphA and replacing it with lacZ,and to understand the regulation of GcvB sRNA to aphA mRNA.The deletion strains were constructed by using λ red homologous recombination,FLP/FRT recombination system and transduction.Subsequently,the corresponding expression changes were analyzed by β-galactosidase activity test and qPCR.The relative transcriptional quantity of aphA decreased in wt,GcvB and its deletion of R1,R2 and R3 as detected by qPCR while no changes of β galactosidase activity were observed for ΔaphA::lacZ,GcvB deletion.When R1,R2 and R3 were absent,the galactosidases activity increased by 244.47%,230.67% and 305.06%,respectively.In conclusion,GcvB might have a certain regulatory effect on aphA.