CAJ | 학술논문

目的研究霍乱弧菌双精氨酸转运系统(twin-arginine translocation system,Tat)基因簇各个基因的共转录情况,并确定参与Tat转录基因簇构成.方法选择霍乱弧菌埃尔托生物型菌株N16961和tatABC基因缺失株N169-dtat进行Tat基因簇及上下游基因,即上游蛋白质生物合成ubi aarF基因和下游细胞色素C551过氧化物酶编码基因cyt551基因进行逆转录,分析Tat基因簇的共转录情况.选择霍乱弧菌及其他弧菌属菌株,通过PCR产物测序和序列比对,对Tat转运系统的同源性进行比较,进行Tat基因簇各基因聚类分析.结果 Tat基因簇4个基因(tatA、tatB、tatC和tatE)均可以共转录.ubi aarF基因可以与tatA及tatB共转录.cyt551基因与上游的4个基因均不共转录;通过N16961 Tat基因缺失株总RNA逆转录分析发现,虽然ubi aarF与tatA、tatB的共转录由于基因缺失被阻断,但该基因仍可以被分别转录.不同型别的霍乱弧菌和其他弧菌属的菌株tatA、tatB、tatC、tatE单基因和tatABC基因簇聚类分析结果表明,Tat基因簇的种属变异性很小,是一段很保守的序列.结论霍乱弧菌ubi aarF基因与Tat基因簇可能共同受调控因子的作用;Tat基因在弧菌属中属于保守结构.
목적 연구곽란호균쌍정안산전운계통(twin-arginine translocation system,Tat)기인족각개기인적공전록정황,병학정삼여Tat전록기인족구성.방법 선택곽란호균애이탁생물형균주N16961화tatABC기인결실주N169-dtat진행Tat기인족급상하유기인,즉상유단백질생물합성ubi aarF기인화하유세포색소C551과양화물매편마기인cyt551기인진행역전록,분석Tat기인족적공전록정황.선택곽란호균급기타호균속균주,통과PCR산물측서화서렬비대,대Tat전운계통적동원성진행비교,진행Tat기인족각기인취류분석.결과 Tat기인족4개기인(tatA、tatB、tatC화tatE)균가이공전록.ubi aarF기인가이여tatA급tatB공전록.cyt551기인여상유적4개기인균불공전록;통과N16961 Tat기인결실주총RNA역전록분석발현,수연ubi aarF여tatA、tatB적공전록유우기인결실피조단,단해기인잉가이피분별전록.불동형별적곽란호균화기타호균속적균주tatA、tatB、tatC、tatE단기인화tatABC기인족취류분석결과표명,Tat기인족적충속변이성흔소,시일단흔보수적서렬.결론 곽란호균ubi aarF기인여Tat기인족가능공동수조공인자적작용;Tat기인재호균속중속우보수결구.
Objective To determine the function of twin-arginine translocation system (Tat) and gene cluster in Vibrio strains and to analyze the homology of tat gene cluster among different Vibrio spp.strains based on N16961 and tatABC mutant strains N169-dtat.Methods Different serotypes of biotype strains of Vibrio spp.were selected to detect the transcription of 4 genes of Tat transport system and upstream ubi aarF gene and downstream cyt551 gene by the total RNA reverse transcription and homologicity of the gene cluster by sequencing analysis.Results Our results showed that the 4 genes of tat cluster (tatA,tatB,tatC,and tatE) were intragenic and co-transcribed.We found that ubi aarF gene could be co-transcribed with tatA,tatB,but not with tatC.The electron transport chain and energy metabolism-related genes,cytochrome C551 peroxidase gene,and 4 genes located at upstream of tatABC operon were not transcribed with tatABC.Althougb the co-transcription between ubi aarF and tatAB was blocked in N169-dtat strain,they were still transcribed separately.Homologous analysis of genes of tat cluster in different types of Vibrio cholerae showed that tat gene cluster was a very conservative.Conclusion The ubi and aarF gene might be co-transcribed with genes of tat cluster in Vibrio cholerae,which and the close relationship showed that they might play a key function in Vibrio cholerae.