烟粉虱内共生菌groEL基因编码一种与植物病毒传毒相关的GroEL蛋白.以B型烟粉虱为研究对象,利用1对特异引物,对B型烟粉虱体内的groEL基因进行PCR扩增,选取502 bp groEL基因片段构建dsRNA,并将其连接植物表达载体pBIN19,将重组质粒电激转...烟粉虱内共生菌groEL基因编码一种与植物病毒传毒相关的GroEL蛋白.以B型烟粉虱为研究对象,利用1对特异引物,对B型烟粉虱体内的groEL基因进行PCR扩增,选取502 bp groEL基因片段构建dsRNA,并将其连接植物表达载体pBIN19,将重组质粒电激转化农杆菌菌株LBA4404.采用农杆菌浸染转化本氏烟草,PCR筛选表明,目的片段已整合至转基因植株的基因组DNA;RT-PCR结果表明,502 bp的groEL基因片段在转基因植株中被转录.展开更多
Objective:To identify members of genera of rickettsia and O.tsutsugammhi simultaneously.Methods:Rapid and duplex and nested PCR methods have been established by designing primers based on the conserved regions of heat...Objective:To identify members of genera of rickettsia and O.tsutsugammhi simultaneously.Methods:Rapid and duplex and nested PCR methods have been established by designing primers based on the conserved regions of heat shock protein GroEL gene.345 mouse viscera samples including liver,spleen and kidney,96 Xenopsylla cheopis and 32 chiggers collected from Hongta areas of Yuxi city,Yunnan province were tested by the new PCR methods.Results:The result of the study showed that the new PCR methods could identify most members of genera -Rickettsia and Orientia simultaneously with 100%specificity and its sensitivity could test one copy per microliter.The results of detection prevalence of rickettsioses in mouse,flea and mites DNA samples showed that the total rickettsia infection rate in mouse was 34.78%(120/345).The total infection rates in R.typhi,O.t Karp and R.sibirica of mouse samples were 28.12%(97/345),19.71%(68/345) and O. 29%(1/345) respectively.Co-infection rates in R.typhi and 0.t Karp of mouse samples were 13.33%(46/ 345).O.t Karp type has been the main epidemic strain in these areas.Conclusion:We concluded that this PCR method could be used to detect multi-genera rickettsia simultaneously.Molecular evidences provided in this and previous studies strongly support that Hongta areas of Yuxi city are a natural focus for typhus and scrub typhus with the common occurrence of their confection.展开更多
Salmonella infection, ranging from mild, self-limiting diarrhea to severe gastrointestinal, septicemic disease and enteric fever, is a global health problem both in humans and animals. Rapid development of microbial d...Salmonella infection, ranging from mild, self-limiting diarrhea to severe gastrointestinal, septicemic disease and enteric fever, is a global health problem both in humans and animals. Rapid development of microbial drug resistance has led to a need for efficacious and affordable vaccines against Salmonella. Microbial heat shock proteins （HSPs）, including HSP60 and HSP70, are the dominant antigens that promote the host immune response. Co-administration of these antigens with cytokines, such as IL-22, which plays an important role in antimicrobial defense, can enhance the immune response and protection against pathogens. Therefore, the aim of the present study was to determine the immunogenicity of rGroEL （Hsp60） of S. Typhi, alone or administered in combination with murine rlL-22, and its protective efficacy against lethal infection with Salmonella, in mice. There was appreciable stimulation of the humoral and cell-mediated immune responses in mice immunized with rGroEL alone. However, co-administration of rGroEL with rlL-22 further boosted the antibody titers （IgG, IgG1 and IgG2a）, T-cell proliferative responses and the secretion of both Thl and Th2 cytokines. Additionally, rGroEL alone accorded 65%-70% protection against lethal challenge with S. Typhi and S. Typhimurium, which increased to 90% when co-administered with rlL-22.展开更多
文摘烟粉虱内共生菌groEL基因编码一种与植物病毒传毒相关的GroEL蛋白.以B型烟粉虱为研究对象,利用1对特异引物,对B型烟粉虱体内的groEL基因进行PCR扩增,选取502 bp groEL基因片段构建dsRNA,并将其连接植物表达载体pBIN19,将重组质粒电激转化农杆菌菌株LBA4404.采用农杆菌浸染转化本氏烟草,PCR筛选表明,目的片段已整合至转基因植株的基因组DNA;RT-PCR结果表明,502 bp的groEL基因片段在转基因植株中被转录.
基金supported by National Natural Science Foundation of China(No. 30771854)China-U.S.Collaborative Program on Emerging and Re-emerging Infectious Diseases(No. 1U2GGH000018-01)
文摘Objective:To identify members of genera of rickettsia and O.tsutsugammhi simultaneously.Methods:Rapid and duplex and nested PCR methods have been established by designing primers based on the conserved regions of heat shock protein GroEL gene.345 mouse viscera samples including liver,spleen and kidney,96 Xenopsylla cheopis and 32 chiggers collected from Hongta areas of Yuxi city,Yunnan province were tested by the new PCR methods.Results:The result of the study showed that the new PCR methods could identify most members of genera -Rickettsia and Orientia simultaneously with 100%specificity and its sensitivity could test one copy per microliter.The results of detection prevalence of rickettsioses in mouse,flea and mites DNA samples showed that the total rickettsia infection rate in mouse was 34.78%(120/345).The total infection rates in R.typhi,O.t Karp and R.sibirica of mouse samples were 28.12%(97/345),19.71%(68/345) and O. 29%(1/345) respectively.Co-infection rates in R.typhi and 0.t Karp of mouse samples were 13.33%(46/ 345).O.t Karp type has been the main epidemic strain in these areas.Conclusion:We concluded that this PCR method could be used to detect multi-genera rickettsia simultaneously.Molecular evidences provided in this and previous studies strongly support that Hongta areas of Yuxi city are a natural focus for typhus and scrub typhus with the common occurrence of their confection.
文摘Salmonella infection, ranging from mild, self-limiting diarrhea to severe gastrointestinal, septicemic disease and enteric fever, is a global health problem both in humans and animals. Rapid development of microbial drug resistance has led to a need for efficacious and affordable vaccines against Salmonella. Microbial heat shock proteins （HSPs）, including HSP60 and HSP70, are the dominant antigens that promote the host immune response. Co-administration of these antigens with cytokines, such as IL-22, which plays an important role in antimicrobial defense, can enhance the immune response and protection against pathogens. Therefore, the aim of the present study was to determine the immunogenicity of rGroEL （Hsp60） of S. Typhi, alone or administered in combination with murine rlL-22, and its protective efficacy against lethal infection with Salmonella, in mice. There was appreciable stimulation of the humoral and cell-mediated immune responses in mice immunized with rGroEL alone. However, co-administration of rGroEL with rlL-22 further boosted the antibody titers （IgG, IgG1 and IgG2a）, T-cell proliferative responses and the secretion of both Thl and Th2 cytokines. Additionally, rGroEL alone accorded 65%-70% protection against lethal challenge with S. Typhi and S. Typhimurium, which increased to 90% when co-administered with rlL-22.
