Clone and functional analysis of gene rpfG in Lysobacter enzymogenes strain OH11 | 产酶溶杆菌rpfG基因的克隆与功能分析
2013
Liu Yiru, Nanjing Agricultural University, Nanjing(China),College of Plant Protection | Xu Feifei, Nanjing Agricultural University, Nanjing(China),College of Plant Protection | Qian Guoliang, Nanjing Agricultural University, Nanjing(China),College of Plant Protection
Chinese. 为探索基因rpfG在产酶溶杆菌中的功能，以产酶溶杆菌OH11为研究对象，在基因组测序的基础上，通过同源比对，存OH11基因组中鉴定出rpfG的同源基因，并命名为rpfGlys。利用同源重组技术，对rpfG基因进行了定向敲除，获得了rpfG的缺失突变株。与野生型相比，rpfG基因的突变降低了产酶溶杆菌重要抗菌物质――热稳定抗菌因子HSAF的产量和对腐霉的拈抗活性。荧光定量PCR显示，在rpfG突变株中负责生物合成HSAF的关键基因Lysegl002651的表达显著下调，与HSAF活性检测结果相吻合。然而，rpfG的突变不改变产酶溶杆菌4种胞外抗菌水解酶的活性。另外，rpfG突变导致产酶溶杆菌菌落表面干燥，但滑行能力提高。
Show more [+] Less [-]English. To analyse the function of rpfG in Lysobacter enzymogenes OH11, a paralogous gene of rpfG was identified through sequence homology analysis and named as rpfGlys. With homologous recombination technology, a deletion mutant of rpfG was generated. Compared with the wild type, the mutant had a low secretion of heat stable antibacterial factor （ HSAF ） which is an important antibacterial substance of Lysobacter enzymogenes and a lower antipathogenic activity toward Pythium aphanidermatum. Fluorogenic quantitative PCR showed that expression of the key gene（ Lysegl002651 ） which is in charge of biosynthesis of HSAF was significantly lower,which coincided with activity detection of HSAF. Further researches found that the mutant did not change the activities of four- type lyric enzymenes of Lysobacter enzymogenes. Moreover, it was of interest that although mutation of rpfG led to acrid surface of colonies, its sliding ability was strengthened.
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