To evaluate the potentiality of applying gene therapy to endotoxemia in high-risk patients, we investigated the effects of transferring an adeno-associated virus serotype 2 （AAV2）-mediated BPI-Fcγ1 gene on protecti...To evaluate the potentiality of applying gene therapy to endotoxemia in high-risk patients, we investigated the effects of transferring an adeno-associated virus serotype 2 （AAV2）-mediated BPI-Fcγ1 gene on protecting mice from challenge of lethal endotoxin. The chimeric BPI-Fcγ1 gene consists of two parts, one encods functional N-terminus （1 to 199 amino acidic residues） of human BPI, which is a bactericidal/permeability-increasing protein, and the other encodes Fc segment of human immunoglobulin G1 （Fcγ1）. Our results indicated that the target protein could be expressed and secreted into the serum of the gene-transferred mice. After lethal endotoxin challenge, the levels of endotoxin and TNF-a in the gene-transferred mice were decreased. The survival rate of the BPI-Fcγ1 gene-transferred mice was markedly increased. Our data suggest that AAV2-mediated chimeric BPI-Fcγ1 gene delivery can potentially be used clinically for the protection and treatment of endotoxemia and endotoxic shock in high-risk individuals. Cellular ＆ Molecular Immunology. 2006;3（3）：221-225.展开更多
An EMS(ethy methanesulfonate)-induced lethal etiolated(le)mutant obtained from the rice variety Zhongjian 100 was characterized by lethal etiolated phenotypes,with significantly reduced levels of chlorophyll a,chlorop...An EMS(ethy methanesulfonate)-induced lethal etiolated(le)mutant obtained from the rice variety Zhongjian 100 was characterized by lethal etiolated phenotypes,with significantly reduced levels of chlorophyll a,chlorophyll b,total chlorophyll,and carotenoids.Additionally,the mutant displayed a significantly decreased number of chloroplast grana,along with irregular and less-stacked grana lamellae.The le mutant showed markedly diminished root length,root surface area,and root volume compared with the wild type.It also exhibited significantly lower catalase activity and total protein content,while peroxidase activity was significantly higher.Using the map-based cloning method,we successfully mapped the LE gene to a 48-kb interval between markers RM16107 and RM16110 on rice chromosome 3.A mutation(from T to C)was identified at nucleotide position 692 bp of LOC_Os03g59640(ChlD),resulting in a change from leucine to proline.By crossing HM133(a pale green mutant with a single-base substitution of A for G in exon 10 of ChlD subunit)with a heterozygous line of le(LEle),we obtained two plant lines heterozygous at both the LE and HM133 loci.Among 15 transgenic plants,3 complementation lines displayed normal leaf color with significantly higher total chlorophyll,chlorophyll a,and chlorophyll b contents.The mutation in le led to a lethal etiolated phenotype,which has not been observed in other ChlD mutants.The mutation in the AAA+domain of ChlD disrupted the interaction of ChlDle with ChlI as demonstrated by a yeast two-hybrid assay,leading to the loss of ChlD function and hindering chlorophyll synthesis and chloroplast development.Consequently,this disruption is responsible for the lethal etiolated phenotype in the mutant.展开更多
In this work asd gene of Shigella flexneri 2a strain T32 was replaced by Vibrio cholerae toxin B subunit (ctxB) gene with DNA recombination in vivo and in vitro. The resulting derivative of T32, designed as FWL01, cou...In this work asd gene of Shigella flexneri 2a strain T32 was replaced by Vibrio cholerae toxin B subunit (ctxB) gene with DNA recombination in vivo and in vitro. The resulting derivative of T32, designed as FWL01, could stably express CtxB, but its growth in LB medium depended on the presence of diaminopimelic acid (DAP). Then form I plasmid of Shigella sonnei strain S7 was labeled with strain T32 asd gene and mobilized into FWL01. Thus a trivalent candidate oral vaccine strain, designed as FSW01, was constructed. In this candidate strain, a balanced-lethal system was constituted between the host strain and the form I plasmid expressing S. sonnei O antigen. Therefore the candidate strain can express stably not only its own O antigen but also CtxB and O antigen of S. sonnei in the absence of any antibiotic. Experiments showed that FSW01 did not invade HeLa cells or cause keratoconjunctivitis in guinea pigs. However, rabbits immunized FSW01 can elicit significant immune responses. In mice and rhesus monkey models, vaccinated animals were protected against the challenges of wild S. flexneri 2a strain 2457T and S. sonnei strain S9.展开更多
Winter rape(Brassica napus L.)is better than other edible oil crops in China,but poor cold resistance is the key factor restricting its development.Hypocotyl length was found closely related to cold tolerance.The corr...Winter rape(Brassica napus L.)is better than other edible oil crops in China,but poor cold resistance is the key factor restricting its development.Hypocotyl length was found closely related to cold tolerance.The correlation between hypocotyl length and semi-lethal low temperature was significant,and the highest correlation between hypocotyl length and LT50 of autumn sowing was 0.9557.When the hypocotyl were treated at low temperature,the cells were seriously damaged and formed cavity structures,with cell walls seriously damaged or merged into each other.The positive regulation gene of hypocotyl length in resistant line of VHTSG 10 was identified as HY5(transcription factor HY5-like)by qPCR,and bZIP transcription factor was found to be its conserved domain.Fused gene by GFP and HY5 from VHTSG 10 was transient transferred into Nicotiana benthamiana cells.Corresponding to the 35S:GFP widely distributed in plasma membrane of leaf epidermis,the fusion protein 35S:HY5-GFP was mainly distributed in nucleus.Thus we regarded BnHY5 gene is a key gene related to cold tolerance and hypocotyl length in B.napus.展开更多
Since the first report of the release of insects carrying a dominant lethal （RIDL） strategy, RIDL strains have been constructed in species including fruit flies and mosquitoes. However, in many insects, identificati...Since the first report of the release of insects carrying a dominant lethal （RIDL） strategy, RIDL strains have been constructed in species including fruit flies and mosquitoes. However, in many insects, identification of sterile and lethal genes needed to generate a RIDL strain is limited by the lack of molecular and genetic information. Here, we created RIDL strains of Drosophila melanogaster using RNA interference （RNAi） of the Pygopus （Pygo） gene, a key component of the Wingless/Wnt signaling pathway. In two transgenic lines, XDll and XD15, we verified lethality in the absence of tetracycline, but we were unable to demonstrate sex-specific lethality. We found that male XD15 adults maintained on medium without tetracycline had a longer lifespan than wild type. This RNAibased RIDL strain may therefore offer the advantages of a transgene that promotes the expression of two contrary actions at different life stages： lethality in larvae and prolonged lifespan in adults, actions that could work together to provide prolonged delivery of lethality by the RIDL system. Use of RNAi can facilitate the development and application of RIDL strategies in a wide range of species.展开更多
Mutants showing spontaneous cell death in the absence of pathogen attack are called lesion mimic mutants （lmm） （Lorrain et al., 2003）. These mutants usually exhibit typical hypersensitive responses （HRs） within ...Mutants showing spontaneous cell death in the absence of pathogen attack are called lesion mimic mutants （lmm） （Lorrain et al., 2003）. These mutants usually exhibit typical hypersensitive responses （HRs） within or around the lesion spots, which are frequently observed in plants challenged with avirulent pathogens （Lorrain et al., 2003）. A number of these mutants have been characterized in rice （Zeng et al., 2004）, Arabidopsis （Guo et al., 2013）, maize （Wang et al., 2013） and barley （Persson et al., 2008）. Most lmm show enhanced resistance to various pathogens （Huang et al., 2011）, because HR is usually accompanied with enhanced defense responses, such as reactive oxygen species （ROS） activation （Qiao et al., 2010） and increased expression of pathogenesis-related genes （Lorrain et al., 2003）. Additionally, most lmm exhibit defects in growth and development due to the disordered physiolog- ical and molecular processes caused by the lesion spots. Thus, lmm are powerful tools for the study of the molecular mech- anisms of cell death, plant development and disease resistance （Lorrain et al., 2003; Babu et al., 2011）.展开更多
文摘To evaluate the potentiality of applying gene therapy to endotoxemia in high-risk patients, we investigated the effects of transferring an adeno-associated virus serotype 2 （AAV2）-mediated BPI-Fcγ1 gene on protecting mice from challenge of lethal endotoxin. The chimeric BPI-Fcγ1 gene consists of two parts, one encods functional N-terminus （1 to 199 amino acidic residues） of human BPI, which is a bactericidal/permeability-increasing protein, and the other encodes Fc segment of human immunoglobulin G1 （Fcγ1）. Our results indicated that the target protein could be expressed and secreted into the serum of the gene-transferred mice. After lethal endotoxin challenge, the levels of endotoxin and TNF-a in the gene-transferred mice were decreased. The survival rate of the BPI-Fcγ1 gene-transferred mice was markedly increased. Our data suggest that AAV2-mediated chimeric BPI-Fcγ1 gene delivery can potentially be used clinically for the protection and treatment of endotoxemia and endotoxic shock in high-risk individuals. Cellular ＆ Molecular Immunology. 2006;3（3）：221-225.
基金supported by the National Natural Science Foundation of China(Grant No.32072049)Open Foundation of State Key Laboratory of Rice Biology,China(Grant No.20210208).
文摘An EMS(ethy methanesulfonate)-induced lethal etiolated(le)mutant obtained from the rice variety Zhongjian 100 was characterized by lethal etiolated phenotypes,with significantly reduced levels of chlorophyll a,chlorophyll b,total chlorophyll,and carotenoids.Additionally,the mutant displayed a significantly decreased number of chloroplast grana,along with irregular and less-stacked grana lamellae.The le mutant showed markedly diminished root length,root surface area,and root volume compared with the wild type.It also exhibited significantly lower catalase activity and total protein content,while peroxidase activity was significantly higher.Using the map-based cloning method,we successfully mapped the LE gene to a 48-kb interval between markers RM16107 and RM16110 on rice chromosome 3.A mutation(from T to C)was identified at nucleotide position 692 bp of LOC_Os03g59640(ChlD),resulting in a change from leucine to proline.By crossing HM133(a pale green mutant with a single-base substitution of A for G in exon 10 of ChlD subunit)with a heterozygous line of le(LEle),we obtained two plant lines heterozygous at both the LE and HM133 loci.Among 15 transgenic plants,3 complementation lines displayed normal leaf color with significantly higher total chlorophyll,chlorophyll a,and chlorophyll b contents.The mutation in le led to a lethal etiolated phenotype,which has not been observed in other ChlD mutants.The mutation in the AAA+domain of ChlD disrupted the interaction of ChlDle with ChlI as demonstrated by a yeast two-hybrid assay,leading to the loss of ChlD function and hindering chlorophyll synthesis and chloroplast development.Consequently,this disruption is responsible for the lethal etiolated phenotype in the mutant.
文摘In this work asd gene of Shigella flexneri 2a strain T32 was replaced by Vibrio cholerae toxin B subunit (ctxB) gene with DNA recombination in vivo and in vitro. The resulting derivative of T32, designed as FWL01, could stably express CtxB, but its growth in LB medium depended on the presence of diaminopimelic acid (DAP). Then form I plasmid of Shigella sonnei strain S7 was labeled with strain T32 asd gene and mobilized into FWL01. Thus a trivalent candidate oral vaccine strain, designed as FSW01, was constructed. In this candidate strain, a balanced-lethal system was constituted between the host strain and the form I plasmid expressing S. sonnei O antigen. Therefore the candidate strain can express stably not only its own O antigen but also CtxB and O antigen of S. sonnei in the absence of any antibiotic. Experiments showed that FSW01 did not invade HeLa cells or cause keratoconjunctivitis in guinea pigs. However, rabbits immunized FSW01 can elicit significant immune responses. In mice and rhesus monkey models, vaccinated animals were protected against the challenges of wild S. flexneri 2a strain 2457T and S. sonnei strain S9.
基金Longdong College youth Science and technology innovation project(XYZK2106)the Agriculture Research System Gansu Province(GARS-TSZ-1)+2 种基金Agriculture Research System of China(CARS-12)Study on variety substitution and key technology of cold resistance and high yield of winter rape(GSLK-2021-12)the National Natural Science Foundation of China(31860388)。
文摘Winter rape(Brassica napus L.)is better than other edible oil crops in China,but poor cold resistance is the key factor restricting its development.Hypocotyl length was found closely related to cold tolerance.The correlation between hypocotyl length and semi-lethal low temperature was significant,and the highest correlation between hypocotyl length and LT50 of autumn sowing was 0.9557.When the hypocotyl were treated at low temperature,the cells were seriously damaged and formed cavity structures,with cell walls seriously damaged or merged into each other.The positive regulation gene of hypocotyl length in resistant line of VHTSG 10 was identified as HY5(transcription factor HY5-like)by qPCR,and bZIP transcription factor was found to be its conserved domain.Fused gene by GFP and HY5 from VHTSG 10 was transient transferred into Nicotiana benthamiana cells.Corresponding to the 35S:GFP widely distributed in plasma membrane of leaf epidermis,the fusion protein 35S:HY5-GFP was mainly distributed in nucleus.Thus we regarded BnHY5 gene is a key gene related to cold tolerance and hypocotyl length in B.napus.
基金supported by the National Natural Science Foundation of China (30800721)
文摘Since the first report of the release of insects carrying a dominant lethal （RIDL） strategy, RIDL strains have been constructed in species including fruit flies and mosquitoes. However, in many insects, identification of sterile and lethal genes needed to generate a RIDL strain is limited by the lack of molecular and genetic information. Here, we created RIDL strains of Drosophila melanogaster using RNA interference （RNAi） of the Pygopus （Pygo） gene, a key component of the Wingless/Wnt signaling pathway. In two transgenic lines, XDll and XD15, we verified lethality in the absence of tetracycline, but we were unable to demonstrate sex-specific lethality. We found that male XD15 adults maintained on medium without tetracycline had a longer lifespan than wild type. This RNAibased RIDL strain may therefore offer the advantages of a transgene that promotes the expression of two contrary actions at different life stages： lethality in larvae and prolonged lifespan in adults, actions that could work together to provide prolonged delivery of lethality by the RIDL system. Use of RNAi can facilitate the development and application of RIDL strategies in a wide range of species.
基金supported by grants from the National Natural Science Foundation of China No. 31401351 to J. Wang, and No. 31171622 and No. 31371705 to X. W. Chensupported by the "Hundred Talents Plan" Foundation of Sichuan to X. Chenthe Specialized Research Fund for Doctoral Program of Higher Education (No. 20135103120004) to J. Wang
文摘Mutants showing spontaneous cell death in the absence of pathogen attack are called lesion mimic mutants （lmm） （Lorrain et al., 2003）. These mutants usually exhibit typical hypersensitive responses （HRs） within or around the lesion spots, which are frequently observed in plants challenged with avirulent pathogens （Lorrain et al., 2003）. A number of these mutants have been characterized in rice （Zeng et al., 2004）, Arabidopsis （Guo et al., 2013）, maize （Wang et al., 2013） and barley （Persson et al., 2008）. Most lmm show enhanced resistance to various pathogens （Huang et al., 2011）, because HR is usually accompanied with enhanced defense responses, such as reactive oxygen species （ROS） activation （Qiao et al., 2010） and increased expression of pathogenesis-related genes （Lorrain et al., 2003）. Additionally, most lmm exhibit defects in growth and development due to the disordered physiolog- ical and molecular processes caused by the lesion spots. Thus, lmm are powerful tools for the study of the molecular mech- anisms of cell death, plant development and disease resistance （Lorrain et al., 2003; Babu et al., 2011）.
