CAJ | 학술논문

为鉴定产甲酸草酸杆菌（Oxalobacter formigenes）是否具有细胞数群体感应（QS）系统，将费氏弧菌（Vibrio fischeri）和荧光假单胞菌（Pseudomonas fluorences）QS系统相关基因与产甲酸草酸杆菌（O. formigenes）基因组序列进行比对，初步确定hdtS和AGPA为QS系统候选基因。通过PCR和T-A克隆方法分别克隆以上两个基因并将其构建成表达载体pET19-hdtS和pET19-AGPA，借助SDS-PAGE凝胶电泳证实以上两个基因均在大肠杆菌（Escherichia coli） BL21（DE3）中高效表达，采用平板菌株报告法鉴定hdtS具有酰基高丝氨酸内酯（AHL）合成酶的功能，而AGPA则不具有该功能。这是国内外首次报道产甲酸草酸杆菌（O. formigenes）中QS系统有关基因，对该菌株QS系统了解及分析影响该菌株在人肠道内定殖因素具有重要意义。
위감정산갑산초산간균（Oxalobacter formigenes）시부구유세포수군체감응（QS）계통，장비씨호균（Vibrio fischeri）화형광가단포균（Pseudomonas fluorences）QS계통상관기인여산갑산초산간균（O. formigenes）기인조서렬진행비대，초보학정hdtS화AGPA위QS계통후선기인。통과PCR화T-A극륭방법분별극륭이상량개기인병장기구건성표체재체pET19-hdtS화pET19-AGPA，차조SDS-PAGE응효전영증실이상량개기인균재대장간균（Escherichia coli） BL21（DE3）중고효표체，채용평판균주보고법감정hdtS구유선기고사안산내지（AHL）합성매적공능，이AGPA칙불구유해공능。저시국내외수차보도산갑산초산간균（O. formigenes）중QS계통유관기인，대해균주QS계통료해급분석영향해균주재인장도내정식인소구유중요의의。
To identify the quorum-sensing system (QS) genes in Oxalobacter formigenes, QS-related genes from Vibrio fischeri and Pseudomonas fluorences were aligned with the genome sequence of O. formigenes. The genes hdtS and AGPA were chosen as the candidate QS-related genes, cloned by PCR into the vector pEASY-T3, and constructed into pET19-pp-histag. The resulting expression vectors designated pET19-hdtS and pET19-AGPA were transformed into Escherichia coli BL21 (DE3), respectively, fol owed by the induction by IPTG. SDS-PAGE showed that the genes hdtS and AGPA were over-expressed in E. coli BL21 (DE3). The hdtS gene, but AGPA not, was final y established to encode an acylhomoserine lactone (AHL) synthase via an AHL reporter strain, Agrobacterium tumefaciens NTIA (pZLR4) through the double-agar diffusion method. To the best of our knowledge, this is the first report on the cloning and identification of the QS-realted gene from O. formigenes, which is very helpful for better understanding of the quorum sensing system of O. formigenes and analysis of impact factors of its colonization in human intestines.