甲型副伤寒沙门菌CMCC 50973的基因表达谱及外膜蛋白BtuB的保护性研究
|
摘要
甲型副伤寒沙门菌（SalmonellaParatyphiA，SPA）是甲型副伤寒（paratyphoidfeverA）的病原菌，通过粪－口途径传播，感染剂量为103～109个菌，能够引起伤寒样临床症状。人是唯一宿主。人群对甲型副伤寒沙门菌普遍易感，但儿童和青壮年发病最高。近些年来，在世界范围内，伤寒发病率有所降低，而甲型副伤寒发病率有明显升高。在我国，甲型副伤寒的发病率也相对较高，并在局部地区有爆发或流行。目前，旧的甲型副伤寒灭活疫苗由于副反应太大，已停止使用，尚无新的有效疫苗预防甲型副伤寒。因此对甲型副伤寒沙门菌的研究也越来越引起肠道致病菌领域学者的重视。
当前分子生物学、基因组学、蛋白组学、生物信息学等快速发展的学科为疫苗的研发提供了革命性手段。通过对基因组信息的分析，筛选候选蛋白质抗原的方法被称为反向疫苗学（reversevaccinology），包括以下主要步骤：在获取病原体的基因组序列后，芯片分析预测编码分泌蛋白、表面相关抗原和毒力因子；蛋白质组学分析膜相关的蛋白；DNA微阵列鉴定高度表达的基因和体内上调的基因；从中综合分析可能的蛋白质抗原，将它们高通量克隆并表达；进行体内或体外免疫学实验分析，筛选有效的候选疫苗。可见，基因组序列和生物信息学分析的快速发展使基因注释不再是限定了总蛋白数和根据预测的功能分成几个组，而是预测细菌的可能全部蛋白在细胞中定位和功能等的信息。此外，蛋白质组学的发展也对疫苗研究起着积极的促进作用。蛋白质组学技术是对基因组学技术的补充，也是疫苗发展的有力技术支撑。综合基因组学、生物信息学、DNA微阵列和蛋白质组学技术，可以对存在于病原体的不同成分给予充分详细的定性、定量测定，对研发亚单位疫苗有很大的帮助。
本研究所使用的甲型副伤寒沙门菌CMCC50973，是一株经筛选的毒力较强的地方流行株，计划用于甲型副伤寒亚单位疫苗的研究。为了全面了解其遗传信息，我们提取了细菌的基因组DNA，委托深圳华大公司测定了CMCC50973的全基因组序列。最终得到1个Scaffold环，全长为4,608,196bp，GC%为52.24%。预测基因4618个，tRNA100种，rRNA7种,sRNA56种。利用KEGG、COG、SwissProt、TrEMBL和NR数据库为基因进行了注释。CMCC50973的基因组通过与已完成测序的甲型副伤寒沙门菌ATCC9150的参考序列NC_006511进行比较，一共找到了143个SNP位点，其中：在基因区的有112个；在基因间区的有31个，共找到12个InDel位点，其中插入位点6个，删除位点6个。该菌全基因组序列的获得，为进一步探寻基因间相互作用、新的调控因子等微生物更详尽的遗传学和生物学信息、预测和筛选新的更特异的保护性抗原基因，并在此基础上发展高效疫苗或经过遗传学操作改造疫苗菌株、构建活疫苗以及发展基因工程菌载体等打下了基础。
表达蛋白质组学的研究是蛋白质组学研究的基础内容。建立起细胞在正常生理条件下的蛋白质参考图谱和数据库，首先能对细菌的蛋白质表达规律有整体的认识，为细菌的研究提供确切、宝贵的数据资料；其次能为后续的功能比较研究，如比较分析在变化了的条件下，蛋白质表达量的变化、翻译后的加工修饰、蛋白质在亚细胞水平上的改变等，提供很好的基础，从而发现和鉴定出特定功能的蛋白及其基因；第三是可以发现和解决一些生物学问题，如找到新的疫苗靶位，也为将来进行病原性研究、疫苗研制（免疫反应性抗原筛选）等相关工作提供重要的参考。因此，我们由地方流行株中筛选出来的毒力较强的菌株CMCC50973，进行了全菌蛋白表达谱的研究。本试验利用pH4.0-5.0、pH4.5-5.5、pH5.0-6.0、pH5.5-6.7的窄梯度胶条和pH6-11的碱性胶条，对CMCC50973对数生长末期的全菌体蛋白进行了双向电泳分离，然后对考马斯亮兰胶上可见的蛋白点取点进行胶内酶切，用MALDI-TOF/TOF-MS鉴定。共取蛋白点848个，鉴定到705个蛋白质点，代表519个基因编码产物，有多个蛋白点鉴定为同一基因产物的现象。所鉴定蛋白通过试验计算得出的分子量和等电点，与基因组注释中的预测值比较一致，但也有一些蛋白质理论值与计算值间存在一些差异。所鉴定蛋白质中多数功能是和能量代谢、营养素的运输和代谢相关；主要出现在糖酵解途径、三羧酸循环、磷酸戊糖途径、嘌呤和嘧啶的代谢等代谢通路中。在试验中鉴定到的67个蛋白被注释为“假想蛋白”（hypotheticalprotein），功能尚不明确。CMCC50973的全菌蛋白质组图谱的获得，帮助我们从整体层面了解了该菌基因组的蛋白表达信息，为研究细菌的致病机理和宿主特异性、基因组的表达情况、寻找新的疫苗靶位、以及免疫反应性抗原筛选等相关工作提供了基础。
外膜蛋白（outermembraneprotein,OMP），是革兰氏阴性菌外膜的主要组成结构，在细菌的黏附、侵袭、耐药性产生等过程中都有着非常重要的作用。因此，在开发该类病原菌的疫苗过程中，对外膜蛋白的研究和筛选，已经成为各类疫苗研发过程中非常重要的内容。我们前期的试验证明甲型副伤寒沙门菌外膜蛋白BtuB与甲型副伤寒恢复期病人的血清有强烈的免疫反应，为了进一步研究该蛋白的免疫保护性，我们在大肠杆菌中克隆、表达了该蛋白，经纯化后获得了纯度约为90%的rBtuB蛋白，小鼠免疫显示重组蛋白具有良好的免疫原性。但对攻毒的保护率仅为60%，可考虑将其用于结合疫苗的载体，与其它抗原共同使用。
Salmonella enterica serovar Paratyphi A (SPA), a Gram-negative bacteria, is an emerging food and water-borne pathogen causing paratyphoid fever and is host-adapted to human only. The World Health Organization (WHO) has estimated that typhoid fever (including paratyphoid fever) causes approximately22million illnesses and220,000deaths per year. SPA is the most prevalent cause of paratyphoid fever. SPA infection rates have been rising, particularly in South East Asia including south of China, where this serovar is now responsible for30-50%of enteric fever cases. This increase has been associated with rises in antibiotic resistance among paratyphoid infections. It may also be associated with vaccination against Typhi, which unfortunately provides little cross-protection against Paratyphi A. Meanwhile, the present use of inactivated whole-cell paratyphoid A vaccines is restricted by their high rates of side effect. It is necessary to research and develop new protective vaccine of paratyphoid A fever. Therefore, the scholars of enteric pathogens field pay more attention to the study of SPA.
At present, the rapid development of molecular biology, genomics, proteomics, bioinformatics and other subjects can provide the revolutionary methods for vaccine research and development. Reverse Vaccinology, an analysis based on the genomic information to screen candidate protein antigens, includes the following main steps: first, after obtaining the pathogen genome sequence, microarray analysis were used to predict encoding secreted proteins, surface-associated antigen and virulence factor; secondly, utility of proteomics to analyze the target proteins, such as outer membrane proteins, or DNA microarrays to identify highly expressed genes and genes regulation in vivo; thirdly, summarize the above results, the candidate protein antigens were defined and were high-throughput cloned and expressed in certain expression system; finally, candidate vaccine were selected by immunological experiments in vivo or in vitro. So, following the rapid development of genomics and bioinformatics analysis, gene annotation is no longer limited to the number of total proteins and function divided into several groups according to the forecast, but can forecast all proteins of cell including more information, such as localization and function. In addition, the development of proteomics plays an active role on vaccine research. Proteomics technology is a complement to genomics technology, also provides effective technical support on vaccine development. Comprehensive application of genomics, bioinformatics, DNA microarray and proteomics technology, can determine different components of the pathogens qualitatively and quantitatively. It is helpful for developing subunit vaccine.
In this study, a reference strain of Salmonella enterica serovar Paratyphi A, CMCC50973, obtained from National Center for Medical Culture Collection (CMCC, Beijing, China), was used. This strain is a stronger virulence isolates from a SPA-infected patient and was planned for the study of paratyphoid fever vaccine. The genomic DNA of CMCC50973strain was extracted and entrusted to BGI-Shenzhen to determine the complete genome sequence. The genome of CMCC50973consists of a single, circular chromosome of4,608,961bp with an average G+C content of52.24%, and including a total of4,618complete open reading frames (ORFs),7rRNA clusters,100tRNAs,56sRNAs. All4,618genes were annotated according to different database of KEGG, COG, SwissProt, TrEMBL, or NR. Comparing with reference NC_006511sequence of SPA strain ATCC9150, there are143single nucleotide polymorphisms (SNPs) and12small insertion or deletion events (Indels). Of the143SNPs,112were located in open reading frames and31were in the intergenic regions. Out of12Indels, insertion events and deletion events have six respectively. In summary, the complete genome sequence of CMCC50973will lay the foundation to explore more detailed microbial genetics and biological information, such as the interaction between genes, the new regulatory factors, prediction and screening of new, more specific protective antigen gene, facilitate additional bioinformation and enable in-depth studies of SPA sequence variations.
Expression proteomics research is the basis of proteomics research. To establish the protein reference map and database of whole-cell proteins under normal physiological conditions, first of all, can help us to have an overall understanding of the expression of protein; second, can provide a good foundation for subsequent comparative proteomics studies, such as comparative analysis the protein expression changes of different conditions of cell survival, post-translational processing and modification, protein changes at the subcellular level, also help for discovery and identification of the proteins with specific function and their genes; third, can help us to find some new vaccine targets, screen immune response antigens, and proceed future pathogenic studies. Therefore, we used the selected virulent SPA strain CMCC50973, performed the whole-cell protein expression profiling studies. In this study, the proteome reference maps of the end of cells growth log phase were thoroughly analyzed by the use of multiple overlapping narrow pH range (pH4.0-5.0, pH4.5-5.5, pH5.0-6.0, pH5.5-6.7, and pH6-11) two-dimensional gel electrophoresis. Altogether,705spots representing519different protein entries were identified by MALDI-TOF/TOF MS. Some gene products were found in multiple spots indicating isoforms. Calculated values of molecular weight and isoelectric point of identified proteins were similar to the predicted values of genome annotation, but there are also some differences. The most identified proteins contain which related to energy production and conversion, transport and metabolism of the three major nutrients, amino acid transport and translation, and so on. We also analyzed the abundance of identified proteins at different stages and confirmed the expression of67hypothetical proteins. CMCC50973whole-cell proteome map can help us understand the genomic expression information from the overall level, and provide a basis for looking for new vaccine targets or screening of the immune response antigen.
Outer membrane proteins (OMPs) of Gram-negative bacteria play an essential role in bacterial pathogenesis and are directly involved in the interaction with host environments. Thus they represent important virulence factors and can induce a good immune response in host, which are usually facilitated the targets of antimicrobial drugs and vaccines. Salmonella OMPs also have been investigated as potential vaccine candidates and diagnostic antigens. Our former test showed that the outer membrane protein BtuB of SPA had a strong immune response with convalescent patient serum of paratyphoid fever. In order to further study the immunogenicity and immune protection of BtuB, the protein was cloned and expressed in Escherichia coli. About90%purity rBtuB protein was obtained after purification. Animal trials have shown that the recombinant protein had good immunogenicity. The lethal challenged experiments showed60%immunized mice with rBtuB were protected. The rBtuB protein showed good immunogenicity but unsatisfactory immune protection, it might be considered for conjugate vaccine carrier and used with other antigens.
当前分子生物学、基因组学、蛋白组学、生物信息学等快速发展的学科为疫苗的研发提供了革命性手段。通过对基因组信息的分析，筛选候选蛋白质抗原的方法被称为反向疫苗学（reversevaccinology），包括以下主要步骤：在获取病原体的基因组序列后，芯片分析预测编码分泌蛋白、表面相关抗原和毒力因子；蛋白质组学分析膜相关的蛋白；DNA微阵列鉴定高度表达的基因和体内上调的基因；从中综合分析可能的蛋白质抗原，将它们高通量克隆并表达；进行体内或体外免疫学实验分析，筛选有效的候选疫苗。可见，基因组序列和生物信息学分析的快速发展使基因注释不再是限定了总蛋白数和根据预测的功能分成几个组，而是预测细菌的可能全部蛋白在细胞中定位和功能等的信息。此外，蛋白质组学的发展也对疫苗研究起着积极的促进作用。蛋白质组学技术是对基因组学技术的补充，也是疫苗发展的有力技术支撑。综合基因组学、生物信息学、DNA微阵列和蛋白质组学技术，可以对存在于病原体的不同成分给予充分详细的定性、定量测定，对研发亚单位疫苗有很大的帮助。
本研究所使用的甲型副伤寒沙门菌CMCC50973，是一株经筛选的毒力较强的地方流行株，计划用于甲型副伤寒亚单位疫苗的研究。为了全面了解其遗传信息，我们提取了细菌的基因组DNA，委托深圳华大公司测定了CMCC50973的全基因组序列。最终得到1个Scaffold环，全长为4,608,196bp，GC%为52.24%。预测基因4618个，tRNA100种，rRNA7种,sRNA56种。利用KEGG、COG、SwissProt、TrEMBL和NR数据库为基因进行了注释。CMCC50973的基因组通过与已完成测序的甲型副伤寒沙门菌ATCC9150的参考序列NC_006511进行比较，一共找到了143个SNP位点，其中：在基因区的有112个；在基因间区的有31个，共找到12个InDel位点，其中插入位点6个，删除位点6个。该菌全基因组序列的获得，为进一步探寻基因间相互作用、新的调控因子等微生物更详尽的遗传学和生物学信息、预测和筛选新的更特异的保护性抗原基因，并在此基础上发展高效疫苗或经过遗传学操作改造疫苗菌株、构建活疫苗以及发展基因工程菌载体等打下了基础。
表达蛋白质组学的研究是蛋白质组学研究的基础内容。建立起细胞在正常生理条件下的蛋白质参考图谱和数据库，首先能对细菌的蛋白质表达规律有整体的认识，为细菌的研究提供确切、宝贵的数据资料；其次能为后续的功能比较研究，如比较分析在变化了的条件下，蛋白质表达量的变化、翻译后的加工修饰、蛋白质在亚细胞水平上的改变等，提供很好的基础，从而发现和鉴定出特定功能的蛋白及其基因；第三是可以发现和解决一些生物学问题，如找到新的疫苗靶位，也为将来进行病原性研究、疫苗研制（免疫反应性抗原筛选）等相关工作提供重要的参考。因此，我们由地方流行株中筛选出来的毒力较强的菌株CMCC50973，进行了全菌蛋白表达谱的研究。本试验利用pH4.0-5.0、pH4.5-5.5、pH5.0-6.0、pH5.5-6.7的窄梯度胶条和pH6-11的碱性胶条，对CMCC50973对数生长末期的全菌体蛋白进行了双向电泳分离，然后对考马斯亮兰胶上可见的蛋白点取点进行胶内酶切，用MALDI-TOF/TOF-MS鉴定。共取蛋白点848个，鉴定到705个蛋白质点，代表519个基因编码产物，有多个蛋白点鉴定为同一基因产物的现象。所鉴定蛋白通过试验计算得出的分子量和等电点，与基因组注释中的预测值比较一致，但也有一些蛋白质理论值与计算值间存在一些差异。所鉴定蛋白质中多数功能是和能量代谢、营养素的运输和代谢相关；主要出现在糖酵解途径、三羧酸循环、磷酸戊糖途径、嘌呤和嘧啶的代谢等代谢通路中。在试验中鉴定到的67个蛋白被注释为“假想蛋白”（hypotheticalprotein），功能尚不明确。CMCC50973的全菌蛋白质组图谱的获得，帮助我们从整体层面了解了该菌基因组的蛋白表达信息，为研究细菌的致病机理和宿主特异性、基因组的表达情况、寻找新的疫苗靶位、以及免疫反应性抗原筛选等相关工作提供了基础。
外膜蛋白（outermembraneprotein,OMP），是革兰氏阴性菌外膜的主要组成结构，在细菌的黏附、侵袭、耐药性产生等过程中都有着非常重要的作用。因此，在开发该类病原菌的疫苗过程中，对外膜蛋白的研究和筛选，已经成为各类疫苗研发过程中非常重要的内容。我们前期的试验证明甲型副伤寒沙门菌外膜蛋白BtuB与甲型副伤寒恢复期病人的血清有强烈的免疫反应，为了进一步研究该蛋白的免疫保护性，我们在大肠杆菌中克隆、表达了该蛋白，经纯化后获得了纯度约为90%的rBtuB蛋白，小鼠免疫显示重组蛋白具有良好的免疫原性。但对攻毒的保护率仅为60%，可考虑将其用于结合疫苗的载体，与其它抗原共同使用。
Salmonella enterica serovar Paratyphi A (SPA), a Gram-negative bacteria, is an emerging food and water-borne pathogen causing paratyphoid fever and is host-adapted to human only. The World Health Organization (WHO) has estimated that typhoid fever (including paratyphoid fever) causes approximately22million illnesses and220,000deaths per year. SPA is the most prevalent cause of paratyphoid fever. SPA infection rates have been rising, particularly in South East Asia including south of China, where this serovar is now responsible for30-50%of enteric fever cases. This increase has been associated with rises in antibiotic resistance among paratyphoid infections. It may also be associated with vaccination against Typhi, which unfortunately provides little cross-protection against Paratyphi A. Meanwhile, the present use of inactivated whole-cell paratyphoid A vaccines is restricted by their high rates of side effect. It is necessary to research and develop new protective vaccine of paratyphoid A fever. Therefore, the scholars of enteric pathogens field pay more attention to the study of SPA.
At present, the rapid development of molecular biology, genomics, proteomics, bioinformatics and other subjects can provide the revolutionary methods for vaccine research and development. Reverse Vaccinology, an analysis based on the genomic information to screen candidate protein antigens, includes the following main steps: first, after obtaining the pathogen genome sequence, microarray analysis were used to predict encoding secreted proteins, surface-associated antigen and virulence factor; secondly, utility of proteomics to analyze the target proteins, such as outer membrane proteins, or DNA microarrays to identify highly expressed genes and genes regulation in vivo; thirdly, summarize the above results, the candidate protein antigens were defined and were high-throughput cloned and expressed in certain expression system; finally, candidate vaccine were selected by immunological experiments in vivo or in vitro. So, following the rapid development of genomics and bioinformatics analysis, gene annotation is no longer limited to the number of total proteins and function divided into several groups according to the forecast, but can forecast all proteins of cell including more information, such as localization and function. In addition, the development of proteomics plays an active role on vaccine research. Proteomics technology is a complement to genomics technology, also provides effective technical support on vaccine development. Comprehensive application of genomics, bioinformatics, DNA microarray and proteomics technology, can determine different components of the pathogens qualitatively and quantitatively. It is helpful for developing subunit vaccine.
In this study, a reference strain of Salmonella enterica serovar Paratyphi A, CMCC50973, obtained from National Center for Medical Culture Collection (CMCC, Beijing, China), was used. This strain is a stronger virulence isolates from a SPA-infected patient and was planned for the study of paratyphoid fever vaccine. The genomic DNA of CMCC50973strain was extracted and entrusted to BGI-Shenzhen to determine the complete genome sequence. The genome of CMCC50973consists of a single, circular chromosome of4,608,961bp with an average G+C content of52.24%, and including a total of4,618complete open reading frames (ORFs),7rRNA clusters,100tRNAs,56sRNAs. All4,618genes were annotated according to different database of KEGG, COG, SwissProt, TrEMBL, or NR. Comparing with reference NC_006511sequence of SPA strain ATCC9150, there are143single nucleotide polymorphisms (SNPs) and12small insertion or deletion events (Indels). Of the143SNPs,112were located in open reading frames and31were in the intergenic regions. Out of12Indels, insertion events and deletion events have six respectively. In summary, the complete genome sequence of CMCC50973will lay the foundation to explore more detailed microbial genetics and biological information, such as the interaction between genes, the new regulatory factors, prediction and screening of new, more specific protective antigen gene, facilitate additional bioinformation and enable in-depth studies of SPA sequence variations.
Expression proteomics research is the basis of proteomics research. To establish the protein reference map and database of whole-cell proteins under normal physiological conditions, first of all, can help us to have an overall understanding of the expression of protein; second, can provide a good foundation for subsequent comparative proteomics studies, such as comparative analysis the protein expression changes of different conditions of cell survival, post-translational processing and modification, protein changes at the subcellular level, also help for discovery and identification of the proteins with specific function and their genes; third, can help us to find some new vaccine targets, screen immune response antigens, and proceed future pathogenic studies. Therefore, we used the selected virulent SPA strain CMCC50973, performed the whole-cell protein expression profiling studies. In this study, the proteome reference maps of the end of cells growth log phase were thoroughly analyzed by the use of multiple overlapping narrow pH range (pH4.0-5.0, pH4.5-5.5, pH5.0-6.0, pH5.5-6.7, and pH6-11) two-dimensional gel electrophoresis. Altogether,705spots representing519different protein entries were identified by MALDI-TOF/TOF MS. Some gene products were found in multiple spots indicating isoforms. Calculated values of molecular weight and isoelectric point of identified proteins were similar to the predicted values of genome annotation, but there are also some differences. The most identified proteins contain which related to energy production and conversion, transport and metabolism of the three major nutrients, amino acid transport and translation, and so on. We also analyzed the abundance of identified proteins at different stages and confirmed the expression of67hypothetical proteins. CMCC50973whole-cell proteome map can help us understand the genomic expression information from the overall level, and provide a basis for looking for new vaccine targets or screening of the immune response antigen.
Outer membrane proteins (OMPs) of Gram-negative bacteria play an essential role in bacterial pathogenesis and are directly involved in the interaction with host environments. Thus they represent important virulence factors and can induce a good immune response in host, which are usually facilitated the targets of antimicrobial drugs and vaccines. Salmonella OMPs also have been investigated as potential vaccine candidates and diagnostic antigens. Our former test showed that the outer membrane protein BtuB of SPA had a strong immune response with convalescent patient serum of paratyphoid fever. In order to further study the immunogenicity and immune protection of BtuB, the protein was cloned and expressed in Escherichia coli. About90%purity rBtuB protein was obtained after purification. Animal trials have shown that the recombinant protein had good immunogenicity. The lethal challenged experiments showed60%immunized mice with rBtuB were protected. The rBtuB protein showed good immunogenicity but unsatisfactory immune protection, it might be considered for conjugate vaccine carrier and used with other antigens.
引文
[1]. Crump, JA; Luby, SP; Mintz, ED, The global burden of typhoid fever. Bull World Health Organ2004,82,(5),346-353.
[2]. DeRoeck, D; Ochiai, RL; Yang, J, et al., Typhoid vaccination:the Asian experience. Expert Rev Vaccines2008,7,(5),547-560.
[3]. Crump, JA; Mintz, ED, Global trends in typhoid and paratyphoid Fever. Clin Infect Dis2010,50,(2),241-246.
[4].魏承毓,我国甲型副伤寒的流行趋势及对防控对策之探讨.国外医学.流行病学传染病学分册2005,32,(2),65-67.
[5].杨进,董柏青,王鸣柳,唐振柱,龚健,李翠云,曾竣,杨宏徽,1994-2002年广西甲型副伤寒和伤寒流行情况分析.中国热带医学2004,4,(2),177-180.
[6].罗芸,叶菊莲,徐宝祥,张政,金大智,占利,杨婷婷,浙江省伤寒副伤寒沙门菌耐药及分子分型.中国公共卫生2010,26,(3),328-330.
[7].姚光海,王涛,田克诚,游旅,韦小俞,唐光鹏,王定明,贵阳市甲型副伤寒沙门菌耐药性分析.预防医学情报杂志2009,25,(8),604-607.
[8].周建芳,杨珊敏,副伤寒甲连续5年流行特点及细菌耐药性监测.中华传染病杂志2003,21,(6),421-422.
[9]. Serruto, D; Serino, L; Masignani, V, et al., Genome-based approaches to develop vaccines against bacterial pathogens. Vaccine2009,27,(25-26),3245-3250.
[10]. Pizza, M; Scarlato, V; Masignani, V, et al., Identification of vaccine candidates against serogroup B meningococcus by whole-genome sequencing. Science2000,287,(5459),1816-1820.
[11]. Wasinger, VC; Cordwell, SJ; Cerpa-Poljak, A, et al., Progress with gene-product mapping of the Mollicutes:Mycoplasma genitalium. Electrophoresis1995,16,(7),1090-1094.
[12]. McClelland, M; Sanderson, KE; Clifton, SW, et al., Comparison of genome degradation in Paratyphi A and Typhi, human-restricted serovars of Salmonella enterica that cause typhoid. Nat Genet2004,36,(12),1268-1274.
[13]. Holt, KE; Thomson, NR; Wain, J, et al., Pseudogene accumulation in the evolutionary histories of Salmonella enterica serovars Paratyphi A and Typhi. BMC Genomics2009,10,36.
[14]. Fleischmann, RD; Adams, MD; White, O, et al., Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science1995,269,(5223),496-512.
[15]. Didelot, X; Achtman, M; Parkhill, J, et al., A bimodal pattern of relatedness between the Salmonella Paratyphi A and Typhi genomes:convergence or divergence by homologous recombination? Genome Res2007,17,(1),61-68.
[16]. Fass, E; Groisman, EA, Control of Salmonella pathogenicity island-2gene expression. Curr Opin Microbiol2009,12,(2),199-204.
[17]. Qin, J; Li, R; Raes, J, et al., A human gut microbial gene catalogue established by metagenomic sequencing. Nature2010,464,(7285),59-65.
[18]. He, F, Human liver proteome project:plan, progress, and perspectives. Mol Cell Proteomics2005,4,(12),1841-1848.
[19]. Wang, J; Huo, K; Ma, L, et al., Toward an understanding of the protein interaction network of the human liver. Mol Syst Biol2011,7,536.
[20]. Li, X; Gong, Y; Wang, Y, et al., Comparison of alternative analytical techniques for the characterisation of the human serum proteome in HUPO Plasma Proteome Project. Proteomics2005,5,(13),3423-3441.
[21]. Gibson, BW; Biemann, K, Strategy for the mass spectrometric verification and correction of the primary structures of proteins deduced from their DNA sequences. Proc Natl Acad Sci U S A1984,81,(7),1956-1960.
[22]. Karas, M; Hillenkamp, F, Laser desorption ionization of proteins with molecular masses exceeding10,000daltons. Anal Chem1988,60,(20),2299-2301.
[23]. Fenn, JB; Mann, M; Meng, CK, et al., Electrospray ionization for mass spectrometry of large biomolecules. Science1989,246,(4926),64-71.
[24]. Romijn, EP; Krijgsveld, J; Heck, AJ, Recent liquid chromatographic-(tandem) mass spectrometric applications in proteomics. J Chromatogr A2003,1000,(1-2),589-608.
[25]. Klose, J, Large-gel2-D electrophoresis. Methods Mol Biol1999,112,147-172.
[26]. Yates, JR,3rd; Link, AJ; Schieltz, D, Direct analysis of proteins in mixtures. Application to protein complexes. Methods Mol Biol2000,146,17-26.
[27]. Washburn, MP; Wolters, D; Yates, JR,3rd, Large-scale analysis of the yeast proteome by multidimensional protein identification technology. Nat Biotechnol2001,19,(3),242-247.
[28]. Washburn, MP; Ulaszek, R; Deciu, C, et al., Analysis of quantitative proteomic data generated via multidimensional protein identification technology. Anal Chem2002,74,(7),1650-1657.
[29]. Gorg, A; Weiss, W; Dunn, MJ, Current two-dimensional electrophoresis technology for proteomics. Proteomics2004,4,(12),3665-3685.
[30]. Yuan, J; Wang, B; Sun, Z, et al., Analysis of host-inducing proteome changes in bifidobacterium longum NCC2705grown in Vivo. J Proteome Res2008,7,(1),375-385.
[31]. Defeu Soufo, HJ; Reimold, C; Linne, U, et al., Bacterial translation elongation factor EF-Tu interacts and colocalizes with actin-like MreB protein. Proc Natl Acad Sci U S A2010,107,(7),3163-3168.
[32]. Balasubramanian, S; Kannan, TR; Baseman, JB, The surface-exposed carboxyl region of Mycoplasma pneumoniae elongation factor Tu interacts with fibronectin. Infect Immun2008,76,(7),3116-3123.
[33]. Castillo-Keller, M; Vuong, P; Misra, R, Novel mechanism of Escherichia coli porin regulation. J Bacteriol2006,188,(2),576-586.
[34]. Li, H; Ye, MZ; Peng, B, et al., Immunoproteomic identification of polyvalent vaccine candidates from Vibrio parahaemolyticus outer membrane proteins. J Proteome Res2010,9,(5),2573-2583.
[35]. Ying, T; Wang, H; Li, M, et al., Immunoproteomics of outer membrane proteins and extracellular proteins of Shigella flexneri2a2457T. Proteomics2005,5,(18),4777-4793.
[36]. Zhu, L; Zhao, G; Stein, R, et al., The proteome of Shigella flexneri2a2457T grown at30and37degrees C. Mol Cell Proteomics2010,9,(6),1209-1220.
[37]. Collins, BS, Gram-negative outer membrane vesicles in vaccine development. Discov Med2011,12,(62),7-15.
[38]. Vogel, U; Claus, H, Vaccine development against Neisseria meningitidis. Microb Biotechnol2011,4,(1),20-31.
[39]. Muralinath, M; Kuehn, MJ; Roland, KL, et al., Immunization with Salmonella enterica serovar Typhimurium-derived outer membrane vesicles delivering the pneumococcal protein PspA confers protection against challenge with Streptococcus pneumoniae. Infect Immun2011,79,(2),887-894.
[40]. Park, SB; Jang, HB; Nho, SW, et al., Outer membrane vesicles as a candidate vaccine against edwardsiellosis. PLoS One2011,6,(3),e17629.
[41]. Parker, H; Chitcholtan, K; Hampton, MB, et al., Uptake of Helicobacter pylori outer membrane vesicles by gastric epithelial cells. Infect Immun2010,78,(12),5054-5061.
[42]. Wilde, H, Enteric fever due to Salmonella typhi and paratyphi A a neglected and emerging problem. Vaccine2007,25,(29),5246-5247.
[43]. Ruan, P; Xia, XP; Sun, D, et al., Recombinant SpaO and Hla as immunogens for protection of mice from lethal infection with Salmonella paratyphi A:implications for rational design of typhoid fever vaccines. Vaccine2008,26,(51),6639-6644.
[44]. Roland, KL; Tinge, SA; Kochi, SK, et al., Reactogenicity and immunogenicity of live attenuated Salmonella enterica serovar Paratyphi A enteric fever vaccine candidates. Vaccine2010,28,(21),3679-3687.
[45]. Gat, O; Galen, JE; Tennant, S, et al., Cell-associated flagella enhance the protection conferred by mucosally-administered attenuated Salmonella Paratyphi A vaccines. PLoS Negl Trop Dis2011,5,(11), e1373.
[46]. Bradbeer, C; Kenley, JS; Di Masi, DR, et al., Transport of vitamin B12in Escherichia coli. Corrinoid specificities of the periplasmic B12-binding protein and of energy-dependent B12transport. J Biol Chem1978,253,(5),1347-1352.
[47]. Zakharov, SD; Eroukova, VY; Rokitskaya, TI, et al., Colicin occlusion of OmpF and TolC channels:outer membrane translocons for colicin import. Biophys J2004,87,(6),3901-3911.
[48]. Gumbart, J; Wiener, MC; Tajkhorshid, E, Coupling of calcium and substrate binding through loop alignment in the outer-membrane transporter BtuB. J Mol Biol2009,393,(5),1129-1142.
[49]. Luan, B; Carr, R; Caffrey, M, et al., The effect of calcium on the conformation of cobalamin transporter BtuB. Proteins2010,78,(5),1153-1162.
[50]. Ochiai, RL; Wang, X; von Seidlein, L, et al., Salmonella paratyphi A rates, Asia. Emerg Infect Dis2005,11,(11),1764-1766.
[51]. Parkhill, J; Dougan, G; James, KD, et al., Complete genome sequence of a multiple drug resistant Salmonella enterica serovar Typhi CT18. Nature2001,413,(6858),848-852.
[52]. Deng, W; Liou, SR; Plunkett, G,3rd, et al., Comparative genomics of Salmonella enterica serovar Typhi strains Ty2and CT18. J Bacteriol2003,185,(7),2330-2337.
[53]. Liu, SL; Sanderson, KE, Rearrangements in the genome of the bacterium Salmonella typhi. Proc Natl Acad Sci U S A1995,92,(4),1018-1022.
[54]. Liu, SL; Sanderson, KE, Homologous recombination between rrn operons rearranges the chromosome in host-specialized species of Salmonella. FEMS Microbiol Lett1998,164,(2),275-281.
[55]. McClelland, M; Sanderson, KE; Spieth, J, et al., Complete genome sequence of Salmonella enterica serovar Typhimurium LT2. Nature2001,413,(6858),852-856.
[56]. Porwollik, S; Frye, J; Florea, LD, et al., A non-redundant microarray of genes for two related bacteria. Nucleic Acids Res2003,31,(7),1869-1876.
[57]. Kingsley, RA; Humphries, AD; Weening, EH, et al., Molecular and phenotypic analysis of the CS54island of Salmonella enterica serotype typhimurium: identification of intestinal colonization and persistence determinants. Infect Immun2003,71,(2),629-640.
[58]. Vernikos, GS; Parkhill, J, Interpolated variable order motifs for identification of horizontally acquired DNA:revisiting the Salmonella pathogenicity islands. Bioinformatics2006,22,(18),2196-2203.
[59]. Blondel, CJ; Jimenez, JC; Contreras, I, et al., Comparative genomic analysis uncovers3novel loci encoding type six secretion systems differentially distributed in Salmonella serotypes. BMC Genomics2009,10,354.
[60]. Lara-Tejero, M; Galan, JE, Salmonella enterica serovar typhimurium pathogenicity island1-encoded type Ⅲ secretion system translocases mediate intimate attachment to nonphagocytic cells. Infect Immun2009,77,(7),2635-2642.
[61]. Ellermeier, CD; Ellermeier, JR; Slauch, JM, HilD, HilC and RtsA constitute a feed forward loop that controls expression of the SPI1type three secretion system regulator hilA in Salmonella enterica serovar Typhimurium. Mol Microbiol2005,57,(3),691-705.
[62]. Daefler, S, Type Ⅲ secretion by Salmonella typhimurium does not require contact with a eukaryotic host. Mol Microbiol1999,31,(1),45-51.
[63]. Abrahams, GL; Hensel, M, Manipulating cellular transport and immune responses:dynamic interactions between intracellular Salmonella enterica and its host cells. Cell Microbiol2006,8,(5),728-737.
[64]. Coombes, BK; Lowden, MJ; Bishop, JL, et al., SseL is a salmonella-specific translocated effector integrated into the SsrB-controlled salmonella pathogenicity island2type Ⅲ secretion system. Infect Immun2007,75,(2),574-580.
[65]. Pickard, D; Wain, J; Baker, S, et al., Composition, acquisition, and distribution of the Vi exopolysaccharide-encoding Salmonella enterica pathogenicity island SPI-7. JBacteriol2003,185,(17),5055-5065.
[66]. Abd E1Ghany, M; Jansen, A; Clare, S, et al., Candidate live, attenuated Salmonella enterica serotype Typhimurium vaccines with reduced fecal shedding are immunogenic and effective oral vaccines. Infect Immun2007,75,(4),1835-1842.
[67]. Kingsley, RA; van Amsterdam, K; Kramer, N, et al., The shdA gene is restricted to serotypes of Salmonella enterica subspecies I and contributes to efficient and prolonged fecal shedding. Infect Immun2000,68,(5),2720-2727.
[68]. Winter, SE; Raffatellu, M; Wilson, RP, et al., The Salmonella enterica serotype Typhi regulator TviA reduces interleukin-8production in intestinal epithelial cells by repressing flagellin secretion. Cell Microbiol2008,10,(1),247-261.
[69]. Baker, S; Holt, K; Whitehead, S, et al., A linear plasmid truncation induces unidirectional flagellar phase change in H:z66positive Salmonella Typhi. Mol Microbiol2007,66,(5),1207-1218.
[70]. Spano, S; Ugalde, JE; Galan, JE, Delivery of a Salmonella Typhi exotoxin from a host intracellular compartment. Cell Host Microbe2008,3,(1),30-38.
[71]. von Rhein, C; Bauer, S; Lopez Sanjurjo, EJ, et al., ClyA cytolysin from Salmonella:distribution within the genus, regulation of expression by SlyA, and pore-forming characteristics. Int J Med Microbiol2009,299,(1),21-35.
[72]. Faucher, SP; Forest, C; Beland, M, et al., A novel PhoP-regulated locus encoding the cytolysin ClyA and the secreted invasin TaiA of Salmonella enterica serovar Typhi is involved in virulence. Microbiology2009,155,(Pt2),477-488.
[73]. Tsui, IS; Yip, CM; Hackett, J, et al., The type IVB pili of Salmonella enterica serovar Typhi bind to the cystic fibrosis transmembrane conductance regulator. Infect Immun2003,71,(10),6049-6050.
[74]. Bishop, A; House, D; Perkins, T, et al., Interaction of Salmonella enterica serovar Typhi with cultured epithelial cells:roles of surface structures in adhesion and invasion. Microbiology2008,154,(Pt7),1914-1926.
[75]. Holt, KE; Parkhill, J; Mazzoni, CJ, et al., High-throughput sequencing provides insights into genome variation and evolution in Salmonella Typhi. Nat Genet2008,40,(8),987-993.
[76]. Wain, J; Diem Nga, LT; Kidgell, C, et al., Molecular analysis of incHIl antimicrobial resistance plasmids from Salmonella serovar Typhi strains associated with typhoid fever. Antimicrob Agents Chemother2003,47,(9),2732-2739.
[77]. Chau, TT; Campbell, JI; Galindo, CM, et al., Antimicrobial drug resistance of Salmonella enterica serovar typhi in asia and molecular mechanism of reduced susceptibility to the fluoroquinolones. Antimicrob Agents Chemother2007,51,(12),4315-4323.
[78]. Holt, KE; Thomson, NR; Wain, J, et al., Multidrug-resistant Salmonella enterica serovar paratyphi A harbors IncHI1plasmids similar to those found in serovar typhi. J Bacteriol2007,189,(11),4257-4264.
[79]. Baker, S; Hardy, J; Sanderson, KE, et al., A novel linear plasmid mediates flagellar variation in Salmonella Typhi. PLoS Pathog2007,3,(5), e59.
[80]. Baker, S; Holt, K; van de Vosse, E, et al., High-throughput genotyping of Salmonella enterica serovar Typhi allowing geographical assignment of haplotypes and pathotypes within an urban District of Jakarta, Indonesia. J Clin Microbiol2008,46,(5),1741-1746.
[81]. Kidgell, C; Pickard, D; Wain, J, et al., Characterisation and distribution of a cryptic Salmonella typhi plasmid pHCM2. Plasmid2002,47,(3),159-171.
[2]. DeRoeck, D; Ochiai, RL; Yang, J, et al., Typhoid vaccination:the Asian experience. Expert Rev Vaccines2008,7,(5),547-560.
[3]. Crump, JA; Mintz, ED, Global trends in typhoid and paratyphoid Fever. Clin Infect Dis2010,50,(2),241-246.
[4].魏承毓,我国甲型副伤寒的流行趋势及对防控对策之探讨.国外医学.流行病学传染病学分册2005,32,(2),65-67.
[5].杨进,董柏青,王鸣柳,唐振柱,龚健,李翠云,曾竣,杨宏徽,1994-2002年广西甲型副伤寒和伤寒流行情况分析.中国热带医学2004,4,(2),177-180.
[6].罗芸,叶菊莲,徐宝祥,张政,金大智,占利,杨婷婷,浙江省伤寒副伤寒沙门菌耐药及分子分型.中国公共卫生2010,26,(3),328-330.
[7].姚光海,王涛,田克诚,游旅,韦小俞,唐光鹏,王定明,贵阳市甲型副伤寒沙门菌耐药性分析.预防医学情报杂志2009,25,(8),604-607.
[8].周建芳,杨珊敏,副伤寒甲连续5年流行特点及细菌耐药性监测.中华传染病杂志2003,21,(6),421-422.
[9]. Serruto, D; Serino, L; Masignani, V, et al., Genome-based approaches to develop vaccines against bacterial pathogens. Vaccine2009,27,(25-26),3245-3250.
[10]. Pizza, M; Scarlato, V; Masignani, V, et al., Identification of vaccine candidates against serogroup B meningococcus by whole-genome sequencing. Science2000,287,(5459),1816-1820.
[11]. Wasinger, VC; Cordwell, SJ; Cerpa-Poljak, A, et al., Progress with gene-product mapping of the Mollicutes:Mycoplasma genitalium. Electrophoresis1995,16,(7),1090-1094.
[12]. McClelland, M; Sanderson, KE; Clifton, SW, et al., Comparison of genome degradation in Paratyphi A and Typhi, human-restricted serovars of Salmonella enterica that cause typhoid. Nat Genet2004,36,(12),1268-1274.
[13]. Holt, KE; Thomson, NR; Wain, J, et al., Pseudogene accumulation in the evolutionary histories of Salmonella enterica serovars Paratyphi A and Typhi. BMC Genomics2009,10,36.
[14]. Fleischmann, RD; Adams, MD; White, O, et al., Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science1995,269,(5223),496-512.
[15]. Didelot, X; Achtman, M; Parkhill, J, et al., A bimodal pattern of relatedness between the Salmonella Paratyphi A and Typhi genomes:convergence or divergence by homologous recombination? Genome Res2007,17,(1),61-68.
[16]. Fass, E; Groisman, EA, Control of Salmonella pathogenicity island-2gene expression. Curr Opin Microbiol2009,12,(2),199-204.
[17]. Qin, J; Li, R; Raes, J, et al., A human gut microbial gene catalogue established by metagenomic sequencing. Nature2010,464,(7285),59-65.
[18]. He, F, Human liver proteome project:plan, progress, and perspectives. Mol Cell Proteomics2005,4,(12),1841-1848.
[19]. Wang, J; Huo, K; Ma, L, et al., Toward an understanding of the protein interaction network of the human liver. Mol Syst Biol2011,7,536.
[20]. Li, X; Gong, Y; Wang, Y, et al., Comparison of alternative analytical techniques for the characterisation of the human serum proteome in HUPO Plasma Proteome Project. Proteomics2005,5,(13),3423-3441.
[21]. Gibson, BW; Biemann, K, Strategy for the mass spectrometric verification and correction of the primary structures of proteins deduced from their DNA sequences. Proc Natl Acad Sci U S A1984,81,(7),1956-1960.
[22]. Karas, M; Hillenkamp, F, Laser desorption ionization of proteins with molecular masses exceeding10,000daltons. Anal Chem1988,60,(20),2299-2301.
[23]. Fenn, JB; Mann, M; Meng, CK, et al., Electrospray ionization for mass spectrometry of large biomolecules. Science1989,246,(4926),64-71.
[24]. Romijn, EP; Krijgsveld, J; Heck, AJ, Recent liquid chromatographic-(tandem) mass spectrometric applications in proteomics. J Chromatogr A2003,1000,(1-2),589-608.
[25]. Klose, J, Large-gel2-D electrophoresis. Methods Mol Biol1999,112,147-172.
[26]. Yates, JR,3rd; Link, AJ; Schieltz, D, Direct analysis of proteins in mixtures. Application to protein complexes. Methods Mol Biol2000,146,17-26.
[27]. Washburn, MP; Wolters, D; Yates, JR,3rd, Large-scale analysis of the yeast proteome by multidimensional protein identification technology. Nat Biotechnol2001,19,(3),242-247.
[28]. Washburn, MP; Ulaszek, R; Deciu, C, et al., Analysis of quantitative proteomic data generated via multidimensional protein identification technology. Anal Chem2002,74,(7),1650-1657.
[29]. Gorg, A; Weiss, W; Dunn, MJ, Current two-dimensional electrophoresis technology for proteomics. Proteomics2004,4,(12),3665-3685.
[30]. Yuan, J; Wang, B; Sun, Z, et al., Analysis of host-inducing proteome changes in bifidobacterium longum NCC2705grown in Vivo. J Proteome Res2008,7,(1),375-385.
[31]. Defeu Soufo, HJ; Reimold, C; Linne, U, et al., Bacterial translation elongation factor EF-Tu interacts and colocalizes with actin-like MreB protein. Proc Natl Acad Sci U S A2010,107,(7),3163-3168.
[32]. Balasubramanian, S; Kannan, TR; Baseman, JB, The surface-exposed carboxyl region of Mycoplasma pneumoniae elongation factor Tu interacts with fibronectin. Infect Immun2008,76,(7),3116-3123.
[33]. Castillo-Keller, M; Vuong, P; Misra, R, Novel mechanism of Escherichia coli porin regulation. J Bacteriol2006,188,(2),576-586.
[34]. Li, H; Ye, MZ; Peng, B, et al., Immunoproteomic identification of polyvalent vaccine candidates from Vibrio parahaemolyticus outer membrane proteins. J Proteome Res2010,9,(5),2573-2583.
[35]. Ying, T; Wang, H; Li, M, et al., Immunoproteomics of outer membrane proteins and extracellular proteins of Shigella flexneri2a2457T. Proteomics2005,5,(18),4777-4793.
[36]. Zhu, L; Zhao, G; Stein, R, et al., The proteome of Shigella flexneri2a2457T grown at30and37degrees C. Mol Cell Proteomics2010,9,(6),1209-1220.
[37]. Collins, BS, Gram-negative outer membrane vesicles in vaccine development. Discov Med2011,12,(62),7-15.
[38]. Vogel, U; Claus, H, Vaccine development against Neisseria meningitidis. Microb Biotechnol2011,4,(1),20-31.
[39]. Muralinath, M; Kuehn, MJ; Roland, KL, et al., Immunization with Salmonella enterica serovar Typhimurium-derived outer membrane vesicles delivering the pneumococcal protein PspA confers protection against challenge with Streptococcus pneumoniae. Infect Immun2011,79,(2),887-894.
[40]. Park, SB; Jang, HB; Nho, SW, et al., Outer membrane vesicles as a candidate vaccine against edwardsiellosis. PLoS One2011,6,(3),e17629.
[41]. Parker, H; Chitcholtan, K; Hampton, MB, et al., Uptake of Helicobacter pylori outer membrane vesicles by gastric epithelial cells. Infect Immun2010,78,(12),5054-5061.
[42]. Wilde, H, Enteric fever due to Salmonella typhi and paratyphi A a neglected and emerging problem. Vaccine2007,25,(29),5246-5247.
[43]. Ruan, P; Xia, XP; Sun, D, et al., Recombinant SpaO and Hla as immunogens for protection of mice from lethal infection with Salmonella paratyphi A:implications for rational design of typhoid fever vaccines. Vaccine2008,26,(51),6639-6644.
[44]. Roland, KL; Tinge, SA; Kochi, SK, et al., Reactogenicity and immunogenicity of live attenuated Salmonella enterica serovar Paratyphi A enteric fever vaccine candidates. Vaccine2010,28,(21),3679-3687.
[45]. Gat, O; Galen, JE; Tennant, S, et al., Cell-associated flagella enhance the protection conferred by mucosally-administered attenuated Salmonella Paratyphi A vaccines. PLoS Negl Trop Dis2011,5,(11), e1373.
[46]. Bradbeer, C; Kenley, JS; Di Masi, DR, et al., Transport of vitamin B12in Escherichia coli. Corrinoid specificities of the periplasmic B12-binding protein and of energy-dependent B12transport. J Biol Chem1978,253,(5),1347-1352.
[47]. Zakharov, SD; Eroukova, VY; Rokitskaya, TI, et al., Colicin occlusion of OmpF and TolC channels:outer membrane translocons for colicin import. Biophys J2004,87,(6),3901-3911.
[48]. Gumbart, J; Wiener, MC; Tajkhorshid, E, Coupling of calcium and substrate binding through loop alignment in the outer-membrane transporter BtuB. J Mol Biol2009,393,(5),1129-1142.
[49]. Luan, B; Carr, R; Caffrey, M, et al., The effect of calcium on the conformation of cobalamin transporter BtuB. Proteins2010,78,(5),1153-1162.
[50]. Ochiai, RL; Wang, X; von Seidlein, L, et al., Salmonella paratyphi A rates, Asia. Emerg Infect Dis2005,11,(11),1764-1766.
[51]. Parkhill, J; Dougan, G; James, KD, et al., Complete genome sequence of a multiple drug resistant Salmonella enterica serovar Typhi CT18. Nature2001,413,(6858),848-852.
[52]. Deng, W; Liou, SR; Plunkett, G,3rd, et al., Comparative genomics of Salmonella enterica serovar Typhi strains Ty2and CT18. J Bacteriol2003,185,(7),2330-2337.
[53]. Liu, SL; Sanderson, KE, Rearrangements in the genome of the bacterium Salmonella typhi. Proc Natl Acad Sci U S A1995,92,(4),1018-1022.
[54]. Liu, SL; Sanderson, KE, Homologous recombination between rrn operons rearranges the chromosome in host-specialized species of Salmonella. FEMS Microbiol Lett1998,164,(2),275-281.
[55]. McClelland, M; Sanderson, KE; Spieth, J, et al., Complete genome sequence of Salmonella enterica serovar Typhimurium LT2. Nature2001,413,(6858),852-856.
[56]. Porwollik, S; Frye, J; Florea, LD, et al., A non-redundant microarray of genes for two related bacteria. Nucleic Acids Res2003,31,(7),1869-1876.
[57]. Kingsley, RA; Humphries, AD; Weening, EH, et al., Molecular and phenotypic analysis of the CS54island of Salmonella enterica serotype typhimurium: identification of intestinal colonization and persistence determinants. Infect Immun2003,71,(2),629-640.
[58]. Vernikos, GS; Parkhill, J, Interpolated variable order motifs for identification of horizontally acquired DNA:revisiting the Salmonella pathogenicity islands. Bioinformatics2006,22,(18),2196-2203.
[59]. Blondel, CJ; Jimenez, JC; Contreras, I, et al., Comparative genomic analysis uncovers3novel loci encoding type six secretion systems differentially distributed in Salmonella serotypes. BMC Genomics2009,10,354.
[60]. Lara-Tejero, M; Galan, JE, Salmonella enterica serovar typhimurium pathogenicity island1-encoded type Ⅲ secretion system translocases mediate intimate attachment to nonphagocytic cells. Infect Immun2009,77,(7),2635-2642.
[61]. Ellermeier, CD; Ellermeier, JR; Slauch, JM, HilD, HilC and RtsA constitute a feed forward loop that controls expression of the SPI1type three secretion system regulator hilA in Salmonella enterica serovar Typhimurium. Mol Microbiol2005,57,(3),691-705.
[62]. Daefler, S, Type Ⅲ secretion by Salmonella typhimurium does not require contact with a eukaryotic host. Mol Microbiol1999,31,(1),45-51.
[63]. Abrahams, GL; Hensel, M, Manipulating cellular transport and immune responses:dynamic interactions between intracellular Salmonella enterica and its host cells. Cell Microbiol2006,8,(5),728-737.
[64]. Coombes, BK; Lowden, MJ; Bishop, JL, et al., SseL is a salmonella-specific translocated effector integrated into the SsrB-controlled salmonella pathogenicity island2type Ⅲ secretion system. Infect Immun2007,75,(2),574-580.
[65]. Pickard, D; Wain, J; Baker, S, et al., Composition, acquisition, and distribution of the Vi exopolysaccharide-encoding Salmonella enterica pathogenicity island SPI-7. JBacteriol2003,185,(17),5055-5065.
[66]. Abd E1Ghany, M; Jansen, A; Clare, S, et al., Candidate live, attenuated Salmonella enterica serotype Typhimurium vaccines with reduced fecal shedding are immunogenic and effective oral vaccines. Infect Immun2007,75,(4),1835-1842.
[67]. Kingsley, RA; van Amsterdam, K; Kramer, N, et al., The shdA gene is restricted to serotypes of Salmonella enterica subspecies I and contributes to efficient and prolonged fecal shedding. Infect Immun2000,68,(5),2720-2727.
[68]. Winter, SE; Raffatellu, M; Wilson, RP, et al., The Salmonella enterica serotype Typhi regulator TviA reduces interleukin-8production in intestinal epithelial cells by repressing flagellin secretion. Cell Microbiol2008,10,(1),247-261.
[69]. Baker, S; Holt, K; Whitehead, S, et al., A linear plasmid truncation induces unidirectional flagellar phase change in H:z66positive Salmonella Typhi. Mol Microbiol2007,66,(5),1207-1218.
[70]. Spano, S; Ugalde, JE; Galan, JE, Delivery of a Salmonella Typhi exotoxin from a host intracellular compartment. Cell Host Microbe2008,3,(1),30-38.
[71]. von Rhein, C; Bauer, S; Lopez Sanjurjo, EJ, et al., ClyA cytolysin from Salmonella:distribution within the genus, regulation of expression by SlyA, and pore-forming characteristics. Int J Med Microbiol2009,299,(1),21-35.
[72]. Faucher, SP; Forest, C; Beland, M, et al., A novel PhoP-regulated locus encoding the cytolysin ClyA and the secreted invasin TaiA of Salmonella enterica serovar Typhi is involved in virulence. Microbiology2009,155,(Pt2),477-488.
[73]. Tsui, IS; Yip, CM; Hackett, J, et al., The type IVB pili of Salmonella enterica serovar Typhi bind to the cystic fibrosis transmembrane conductance regulator. Infect Immun2003,71,(10),6049-6050.
[74]. Bishop, A; House, D; Perkins, T, et al., Interaction of Salmonella enterica serovar Typhi with cultured epithelial cells:roles of surface structures in adhesion and invasion. Microbiology2008,154,(Pt7),1914-1926.
[75]. Holt, KE; Parkhill, J; Mazzoni, CJ, et al., High-throughput sequencing provides insights into genome variation and evolution in Salmonella Typhi. Nat Genet2008,40,(8),987-993.
[76]. Wain, J; Diem Nga, LT; Kidgell, C, et al., Molecular analysis of incHIl antimicrobial resistance plasmids from Salmonella serovar Typhi strains associated with typhoid fever. Antimicrob Agents Chemother2003,47,(9),2732-2739.
[77]. Chau, TT; Campbell, JI; Galindo, CM, et al., Antimicrobial drug resistance of Salmonella enterica serovar typhi in asia and molecular mechanism of reduced susceptibility to the fluoroquinolones. Antimicrob Agents Chemother2007,51,(12),4315-4323.
[78]. Holt, KE; Thomson, NR; Wain, J, et al., Multidrug-resistant Salmonella enterica serovar paratyphi A harbors IncHI1plasmids similar to those found in serovar typhi. J Bacteriol2007,189,(11),4257-4264.
[79]. Baker, S; Hardy, J; Sanderson, KE, et al., A novel linear plasmid mediates flagellar variation in Salmonella Typhi. PLoS Pathog2007,3,(5), e59.
[80]. Baker, S; Holt, K; van de Vosse, E, et al., High-throughput genotyping of Salmonella enterica serovar Typhi allowing geographical assignment of haplotypes and pathotypes within an urban District of Jakarta, Indonesia. J Clin Microbiol2008,46,(5),1741-1746.
[81]. Kidgell, C; Pickard, D; Wain, J, et al., Characterisation and distribution of a cryptic Salmonella typhi plasmid pHCM2. Plasmid2002,47,(3),159-171.