CAJ | 학술논문

对球形芽孢杆菌(Lysiniacillus sphaericus)C3-41在不同铵盐浓度中的生长进行了研究. 结果表明,在铵盐浓度为0.5~4.0 g/L范围内时,菌株正常生长并形成毒素,但是芽孢形成时间随铵盐浓度的升高而提前. 从该菌株中提取基因组DNA,通过PCR的方法克隆了其基因组中疑似铵转运蛋白质的编码基因amt基因序列 (GeneID: 501248970). 为了研究amt基因在T7启动子启动下的表达, 构建了表达质粒pETAMT并转化进大肠杆菌中,对得到的重组菌株进行活性分析. 结果表明,重组菌株E-pETAMT与对照菌株E-pET28a相比具有明显的将铵盐转运进细胞内及将铵盐分泌到细胞外的活性,即表明该amt基因编码的蛋白质在重组菌株E-pETAMT中表达并表现出铵转运蛋白质的活性. 通过KEGG软件对C3-41菌株氮代谢途径的分析表明,铵盐既用于其氨基酸的合成,也是部分氨基酸的代谢产物.
대구형아포간균(Lysiniacillus sphaericus)C3-41재불동안염농도중적생장진행료연구. 결과표명,재안염농도위0.5~4.0 g/L범위내시,균주정상생장병형성독소,단시아포형성시간수안염농도적승고이제전. 종해균주중제취기인조DNA,통과PCR적방법극륭료기기인조중의사안전운단백질적편마기인amt기인서렬 (GeneID: 501248970). 위료연구amt기인재T7계동자계동하적표체, 구건료표체질립pETAMT병전화진대장간균중,대득도적중조균주진행활성분석. 결과표명,중조균주E-pETAMT여대조균주E-pET28a상비구유명현적장안염전운진세포내급장안염분비도세포외적활성,즉표명해amt기인편마적단백질재중조균주E-pETAMT중표체병표현출안전운단백질적활성. 통과KEGG연건대C3-41균주담대사도경적분석표명,안염기용우기안기산적합성,야시부분안기산적대사산물.
The effect of ammonium on Lysiniacillus sphaericus C3-41 strain was studied, and the result showed that, at the concentration of 0.5~4.0 g/L,C3-41 strain grew in similar level and expressed binary toxins normally, but spores formed earlier at high concentration.The putative ammonium transporter encoding gene (amt, GeneID: 501248970) was cloned by PCR from Lysinibacillus sphaericus C3-41 genomic DNA. To express amt gene under T7 promoter (a vegetative promoter),the recombi-nant plasmid pETAMT was constructed and transformed into Escherichia coli BL21 strain. The generated recombinant strain E-pETAMT could transport ammonium into the cell and secrete ammonium to the extracellular. This result indicated that the pro-tein encoding by amt gene had the activity of ammonium transporter. KEGG analysis of nitrogen metabolism for strain C3-41showed that ammonium could be used for amino acid synthesis,but also was the metabolite product of some amino acids.