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The isopentenol utilization pathway (IUP) is potential in terpenoids synthesis. This study aimed to construct IUP-employed Escherichia coli chassis for stably synthesizing terpenoids. As to effectiveness, promotor engineering strategy was employed to regulate IUP expression level, while ribosome-binding site (RBS) library of the key enzyme was constructed for screening the optimal RBS, followed by optimization of concentration of inducer and substrates, the titer of reporting production, lycopene, from 0.087 to 8.67 mg OD600 -1 . As about stability, the IUP expression cassette was integrated into the genome through transposition tool based on CRISPR-associated transposases. Results showed that the strain with 13 copies produced 1.78-fold lycopene titer that of the controlled strain with IUP-harbored plasmid, and it exhibited stable expression after ten successions while the plasmid loss was observed in the controlled strain in the 3rd succession. This strategy provides valuable information for rapid construction of highly effective and stable chassis employing IUP for terpenoids production.
Asunto(s)
Escherichia coli , Terpenos , Escherichia coli/genética , Escherichia coli/metabolismo , Terpenos/metabolismo , Licopeno/metabolismo , Pentanoles/metabolismo , Ingeniería MetabólicaRESUMEN
Objective To understand the epidemic characteristics of influenza in Minhang District of Shanghai during 2013-2016,so as to provide scientific basis for the prevention and control of influenza.Methods Surveillance data of influenza like illness (ILI) cases from the national influenza network and data of pathogen detection during 2013-2016 in Minhang District were selected to analyze the epidemic characteristics of influenza.Results The number of influenza like illness (ILI) was 37 738 from 2013 to 2016,which accounted for 0.90％ of the outpatients in monthly average.Meanwhile,4 094 samples were collected,among which 857 samples were virus positive.The total isolation rate was 20.93％.In 2013,the dominant strain was mainly seasonal influenza A (H3N2).Dominant strains of 2014 were influenza B (Yamagata) and seasonal influenza A (H3N2).In 2015,dominant strain was seasonal influenza A (H3N2) of summer peak,but the dominant strains of winter-spring peak were new type influenza H1N1 (H1N1 pdm09) and influenza B (Victoria) from December 2015 to April 2016.The dominant strain of winter peak in 2016 was seasonal influenza A (H3N2) popular.Spearman correlation analysis showed positive rate between ILI visiting rate (ILI％) and influenza virus (rs =0.460,P＜0.05).Conclusions The epidemic peaks of of influenza were winter-spring and summer in Minhang District,advantages strains were interaction pandemic with certain regulation.The characteristics of ILI visiting rate was coincident with influenza virus activity.
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Objective To investigate the epidemiological and pathogenic characteristics of viral diarrhea in Minhang District of Shanghai.Methods Random sampling on diarrhea was conducted in intestinal outpatient departments of 2 sentinel hospitals according to a certain sampling interval in Minhang District from 2014 to 2016.Real-time PCR was used to detect rotavirus (RV),norovirus (NV),adenovirus (AD),astrovirus (AstV) and sapovirus (SaV) in fecal samples.Results A total of 11 243 cases of diarrhea were monitored in 2 sentinel hospitals during 2014 and 2016,with 3 213,3 600 and 4 430 cases for each year,respectively.Out of 809 stool specimens,309 were tested positively,and the positive rate was 38.48％.All 5 pathogenic viruses were detected,mostly NV (207 cases,66.99％)followed by RV (77 cases,24.92％).Conclusions NV accounted for the majority of reported infection diarrhea cases in Minhang District of Shanghai from 2014 to 2016,with significant seasonal peaks.Tailored prevention and control measures should be carried out,particularly in risk seasons.
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Objective To investigate the etiological agents of the outbreak of hand,foot and mouth disease (HFMD) in Minhang District of Shanghai from 2009 to 2016,and to provide evidence for the prevention and management policy of HFMD by collecting suspected HFMD samples for laboratory testing from HFMD sentinel hospitals,the Children's Hospitals of Fudan University and jurisdiction community health service centers.Methods Specimens including stools,throat swabs and anal swabs were collected from patients suspected of HFMD from surveillance hospitals,i.e.,Children's Hospital of Fudan University and jurisdiction community health service centers during 2009 and 2016.The specimens were detected by real-time RT-PCR with the five types of viruses including panenterovirus (EV),enterovirus 71 (EV71),Coxsackie virus A16 (CVA16),Coxsackie virus A6 (CVA6) and Coxsackie virus A10 (CVA10).The distribution characteristics of pathogens were analyzed.Results During 2009 and 2016 we collected 3 744 cases of HFMD,the positive detection rate of pan-enterovirus were 84.83％ (3 176 cases),including EV71 (64.45％) and CVA16 (15.77％),then were CVA6 (9.23％) and other EV (8.78％),and CVA10 was only 0.76％.The major etiological agent was EV71.The advantage of different years and different season strain presented dynamic change,mainly EV71 and CVA16 co-popular in 2009,mainly EV71 epidemic from 2010 to 2011,EV71 and CVA16 were co-popular again in 2012,and the priority was EV71,then EV71 and other EV were co-popular in 2013.After 2014,CVA6 gradually increased,mainly CVA6 was popular 2015 and 2016,especially in 2016,the proportion of CVA6 ranged as high as 40.55％.CVA10 showed sporadic distribution trend in very low proportion.From the difference of age and gender in HFMD cases,the males were more than the females (1.68 ∶ 1),and the incidence was the highest in children of 1-3 years old.The peak of incidence occured in April to July,then in September to November.Conclusions HFMD onset seasons mainly were summer and fall-and-winter in bimodal popular models.It happens in children under 5 years of age.The incidence was higher in male than in female.The advantage of different years and different season strains present dynamic change,and the prevalence has obvious age and season limit.
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Objective To understand the epidemic characteristics of influenza in Minhang District of Shanghai during 2013-2016,so as to provide scientific basis for the prevention and control of influenza.Methods Surveillance data of influenza like illness (ILI) cases from the national influenza network and data of pathogen detection during 2013-2016 in Minhang District were selected to analyze the epidemic characteristics of influenza.Results The number of influenza like illness (ILI) was 37 738 from 2013 to 2016,which accounted for 0.90％ of the outpatients in monthly average.Meanwhile,4 094 samples were collected,among which 857 samples were virus positive.The total isolation rate was 20.93％.In 2013,the dominant strain was mainly seasonal influenza A (H3N2).Dominant strains of 2014 were influenza B (Yamagata) and seasonal influenza A (H3N2).In 2015,dominant strain was seasonal influenza A (H3N2) of summer peak,but the dominant strains of winter-spring peak were new type influenza H1N1 (H1N1 pdm09) and influenza B (Victoria) from December 2015 to April 2016.The dominant strain of winter peak in 2016 was seasonal influenza A (H3N2) popular.Spearman correlation analysis showed positive rate between ILI visiting rate (ILI％) and influenza virus (rs =0.460,P＜0.05).Conclusions The epidemic peaks of of influenza were winter-spring and summer in Minhang District,advantages strains were interaction pandemic with certain regulation.The characteristics of ILI visiting rate was coincident with influenza virus activity.
RESUMEN
Objective To investigate the epidemiological and pathogenic characteristics of viral diarrhea in Minhang District of Shanghai.Methods Random sampling on diarrhea was conducted in intestinal outpatient departments of 2 sentinel hospitals according to a certain sampling interval in Minhang District from 2014 to 2016.Real-time PCR was used to detect rotavirus (RV),norovirus (NV),adenovirus (AD),astrovirus (AstV) and sapovirus (SaV) in fecal samples.Results A total of 11 243 cases of diarrhea were monitored in 2 sentinel hospitals during 2014 and 2016,with 3 213,3 600 and 4 430 cases for each year,respectively.Out of 809 stool specimens,309 were tested positively,and the positive rate was 38.48％.All 5 pathogenic viruses were detected,mostly NV (207 cases,66.99％)followed by RV (77 cases,24.92％).Conclusions NV accounted for the majority of reported infection diarrhea cases in Minhang District of Shanghai from 2014 to 2016,with significant seasonal peaks.Tailored prevention and control measures should be carried out,particularly in risk seasons.
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Objective To investigate the etiological agents of the outbreak of hand,foot and mouth disease (HFMD) in Minhang District of Shanghai from 2009 to 2016,and to provide evidence for the prevention and management policy of HFMD by collecting suspected HFMD samples for laboratory testing from HFMD sentinel hospitals,the Children's Hospitals of Fudan University and jurisdiction community health service centers.Methods Specimens including stools,throat swabs and anal swabs were collected from patients suspected of HFMD from surveillance hospitals,i.e.,Children's Hospital of Fudan University and jurisdiction community health service centers during 2009 and 2016.The specimens were detected by real-time RT-PCR with the five types of viruses including panenterovirus (EV),enterovirus 71 (EV71),Coxsackie virus A16 (CVA16),Coxsackie virus A6 (CVA6) and Coxsackie virus A10 (CVA10).The distribution characteristics of pathogens were analyzed.Results During 2009 and 2016 we collected 3 744 cases of HFMD,the positive detection rate of pan-enterovirus were 84.83％ (3 176 cases),including EV71 (64.45％) and CVA16 (15.77％),then were CVA6 (9.23％) and other EV (8.78％),and CVA10 was only 0.76％.The major etiological agent was EV71.The advantage of different years and different season strain presented dynamic change,mainly EV71 and CVA16 co-popular in 2009,mainly EV71 epidemic from 2010 to 2011,EV71 and CVA16 were co-popular again in 2012,and the priority was EV71,then EV71 and other EV were co-popular in 2013.After 2014,CVA6 gradually increased,mainly CVA6 was popular 2015 and 2016,especially in 2016,the proportion of CVA6 ranged as high as 40.55％.CVA10 showed sporadic distribution trend in very low proportion.From the difference of age and gender in HFMD cases,the males were more than the females (1.68 ∶ 1),and the incidence was the highest in children of 1-3 years old.The peak of incidence occured in April to July,then in September to November.Conclusions HFMD onset seasons mainly were summer and fall-and-winter in bimodal popular models.It happens in children under 5 years of age.The incidence was higher in male than in female.The advantage of different years and different season strains present dynamic change,and the prevalence has obvious age and season limit.
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OBJECTIVE: Response surface analysis methodology (RSM) was used to optimize the extraction technology of total flavonoids from Mulgedium tataricum. METHODS: Ethanol concentration, solvent-solid ratio, extraction temperature and extraction time were selected as influencing factors during extraction. The experiment mathematical model was arranged according to Box-Behnken central composite experiment design. RESULTS: The optimal extraction conditions were as following: ethanol concentration 63.78%, solvent-solid ratio 40: 1, extraction temperature 90 degrees C, and extraction time 1.07 h, the predicted value of extraction rate of total flavonoids was 14.04% under the optimum conditions, and the experimental value was 13.79%. CONCLUSION: The combination of Box-Behnken design and response surface analysis can well optimize the extraction technology of total flavonoids from Mulgedium tataricum.