cord-004592-a3k56ulv	2012	To prepare soluble and bioactive recombinant human FGF23 to meet the increasing demand in its pharmacological application, small ubiquitin-related modifier (SUMO)-FGF23 fusion gene and FGF23 non-fusion gene were amplified by standard PCR methods and cloned into vector pET-22b and pET-3c, then transformed into Escherichia coli Rosetta (DE3) and BL21 (DE3). An analysis was performed to determine the effects of the three factors (IPTG concentration, temperature, and time after induction) on the expression yield and productivity of soluble FGF23 in Rosetta(DE3)/pET22b-SUMO-FGF23. The results showed that, except for the combination of pET22b-SUMO-FGF23 and pET3c-SUMO-FGF23, the two kinds of plasmid, and the Rosetta(DE3) host strain (Fig. 3a) , all other combinations had no target protein expressed. Concerning optimal cell growth conditions for protein production, we screened for suitable IPTG concentration, temperature, and time after induction for the expression yield and productivity of soluble FGF23 in Rosetta(DE3)/ pET22b/SUMO-FGF23. High-level expression and purification of soluble recombinant FGF21 protein by SUMO fusion in Escherichia coli
cord-010914-08omkszy	2020	
cord-010991-fp8hljbq	2020	For vaccine development, attention was paid on the purified antigens involved in the pathogenesis of dental caries for the development of potentially safer vaccines, which may reduce the viability of bacteria in the saliva, impairing the surface adhesion and inhibiting the metabolically active enzymes involved in caries formation (Chen and Wang 2010) . mutans have shown encouraging results related to dental caries protection, but were limited by the cross-reactive epitopes against human heart and skeleton muscle tissues as detected by indirect immunofluorescence and crossed immunoelectrophoresis (Kt et al. In the present study, we have tried to evaluate the effect of anti-dextransucrase antibodies on caries formation by using purified dextransucrase as the antigen from S. Protein samples of the liver, heart, spleen, kidney of mice, rat and liver, kidney of rabbit were analysed by western blot analysis to evaluate the cross reactivity of purified antibody with mammalian tissues Fig. 5a .
cord-011012-5mev3otu	2020	It was observed by ELISA that Fubc fusion protein elicited higher serum IgG antibody response either in the presence or in absence of Freund''s adjuvant in comparison to the immune response of Fu and bc peptides separately. Herein, we have expressed the recombinant Fubc antigenic protein, optimised its large-scale purification protocol and finally evaluated its protein-specific immune response in BALB/c mice. Therefore, these two conserved Fu and bc loops have been selected to design a recombinant Fubc antigenic protein for the development of dengue subunit vaccine. Immune response to the purified Fubc protein in BALB/c mice From indirect ELISA, it was observed that the serum IgG levels in both male and female groups treated with Fubc proteins were significantly higher than those treated with only adjuvant and PBS control (Fig. 4a, b) . Antibodies to envelope glycoprotein of dengue virus during the natural course of infection are predominantly cross-reactive and recognize epitopes containing highly conserved residues at the fusion loop of domain II
cord-011147-55whf8md	2020	title: Oral delivery of Bacillus subtilis spores expressing Clonorchis sinensis paramyosin protects grass carp from cercaria infection Potential pathogenetic bacteria with lower abundance and higher abundances of candidate probiotics and bacteria associated with digestion in 1 × 10(11) CFU/g B.s-CotC-CsPmy spores administrated fishes could be detected compared with control group. In the present study, the specific IgM antibody levels in serum, bile, intestinal mucus, and skin mucus of grass carp orally administrated with different dosages of spores (B.s-CotC-CsPmy) were significantly increased with dosedependent from the 2nd week after the beginning of the immunization till to 6 weeks (Fig. 2) . Immune response induced by oral delivery of Bacillus subtilis spores expressing enolase of Clonorchis sinensis in grass carps (Ctenopharyngodon idellus)
cord-011212-ovjdzyxv	2019	
cord-011320-cwvkox29	2020	
cord-012054-bpgb7tgo	2008	
cord-012056-8b3xffsh	2008	
cord-012058-ds7u3ke9	2008	
cord-012062-xlw92xoe	2008	
cord-012064-egzl6zk9	2009	The hps and phi genes of Bacillus brevis, encoding two key steps of the ribulose monophosphate (RuMP) pathway, were introduced to construct a pathway for the metabolism of the toxic methanol oxidation intermediate formaldehyde. Black arrows indicate the assimilatory RuMP pathway, dashed arrows indicate the dissimilatory RuMP pathway, gray arrows indicate the linear oxidation of formaldehyde to carbon dioxide formate dehydrogenase (Fmd) from the methylotrophic yeast Hansenula polymorpha in Saccharomyces cerevisiae resulted in an increased biomass yield with formaldehyde as auxiliary substrate (Baerends et al. The RuMP pathway strain showed significantly improved performance over the control strain, achieving a biomass yield-on-glucose of 91% when using formaldehyde as auxiliary substrate. putida S12pJNNhp(t) and an empty vector control strain were cultured in a C-limited chemostat with glucose as the primary carbon source and formaldehyde as auxiliary substrate. b Biomass yield increase (on glucose) as function of relative formaldehyde concentration in chemostat cultures of strains S12pJNNhp(t) (triangles) and S12pJNN (t) (squares).
cord-012066-8anhcyia	2009	
cord-012072-98wuq5ri	2009	
cord-012085-ubdzhkfq	2011	The diversity and abundance of ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) in the sediment of the Pearl River Estuary were investigated by cloning and quantitative real-time polymerase chain reaction (qPCR). From one sediment sample S16, 36 AOA OTUs (3% cutoff) were obtained from three clone libraries constructed using three primer sets for amoA gene. The terminal restriction fragment length polymorphism results showed that the relative abundance of AOB species in the sediment samples of different salinity were significantly different, indicating that salinity might be a key factor shaping the AOB community composition. Overall, AOB amoA gene copy number was 100 times lower than that of AOA, suggesting that the dominant ammonia-oxidizing microorganisms in the sediment of the Pearl River Estuary were AOA.
cord-012086-sqv56mmq	2011	The key precursors for p-hydroxybenzoate production by engineered Pseudomonas putida S12 are phosphoenolpyruvate (PEP) and erythrose-4-phosphate (E4P), for which the pentose phosphate (PP) pathway is an important source. Even without xylose-co-feeding, p-hydroxybenzoate production was improved in the evolved xylose-utilizing strain, which may indicate an intrinsically elevated PP pathway activity. Pseudomonas putida S12 is a solvent-tolerant bacterium that has been developed as a platform host for the production of a range of substituted aromatic compounds such as phenol, tcinnamate, p-coumarate, p-hydroxybenzoate, and p-hydroxystyrene (Nijkamp et al. putida S12pal_xylB7 on glucose as single carbon source showed a product-tosubstrate yield of 4.9 Cmol%, with a specific production (Fig. 3b) . Because of the improved biomass yield, the effect of the primary substrate on the specific phydroxybenzoate production rate q p was limited: the q p on glycerol (8.57 μmol C (g CDW) −1 h −1 ) was only slightly higher than on glucose (7.96 μmol C (g CDW) −1 h −1 ).
cord-012089-haqqrwad	2011	Since the feasibility of a bioprocess depends critically on the final product titer (Stephanopoulos 2007) , a synthetic approach to produce higher alcohols from nonfermentative pathways in Escherichia coli was previously devised (Atsumi et al. 2008) , in the present work, gas stripping integrated with fermentation was used to compare the isobutanol production by the high producer (JCL260), isobutanol-tolerant (SA481), and the parental (JCL16) strains carrying pSA65 and pSA69 plasmids. Error bars represent the difference between duplicate cultures Fig. 4 Time profile of acetate production by JCL260 PoxB − mutant and high isobutanol producer (JCL260) strains harboring pSA65/pSA69 plasmids cultivated in shake flasks containing 25 mL of medium. (2010b) found that isobutanol production, cell growth, and glucose consumption rate of SA481 tested in flask cultures were similar to those of JCL260 and that improved tolerance of SA481 did not increase the final titer.
cord-012090-pnt4y7zd	2011	
cord-012095-vzk2m91v	2011	
cord-012096-d7e89x03	2011	
cord-012098-1nxws0dk	2011	
cord-012099-fveq5c9w	2011	
cord-012101-mfgca1ou	2011	
cord-012430-3uvhoca9	2008	
cord-012431-l2i5utne	2008	title: Microbiological analysis of the population of extremely haloalkaliphilic sulfur-oxidizing bacteria dominating in lab-scale sulfide-removing bioreactors Therefore, the feasibility of sulfide oxidation into elemental sulfur under oxygen limitation was tested at extremely haloalkaline conditions in lab-scale bioreactors using mix sediments from hypersaline soda lakes as inoculum. The microbiological analysis, both culture dependent and independent, of the successfully operating bioreactors revealed a domination of obligately chemolithoautotrophic and extremely haloalkaliphilic sulfur-oxidizing bacteria belonging to the genus Thioalkalivibrio. Molecular analysis of the biomass from two SL-BR and two FBR bioreactors based on 16S rRNA gene DGGE showed low genetic diversity, typical for autotrophic mix cultures with domination of one to two SOB genotypes and some side heterotrophic populations (Fig. 4) . In the natural soda lake sediments, often another genus of low salt-tolerant alkaliphilic SOB (Thioalkalimicrobium) can be found, which is characterized by extremely high rates of sulfide oxidation (Sorokin and Kuenen 2005; Sorokin et al.
cord-012432-te3sysra	2008	
cord-012435-tt44dkqd	2008	
cord-012437-2r6egrca	2009	
cord-012439-28vi4c2j	2008	
cord-012448-9cuq0jqg	2008	
cord-026729-hn0q0sbv	2020	
cord-257136-zpeh8pmc	2019	To develop a simple, rapid, accurate, and high-throughput detection method for diagnosis and differential diagnosis on swine enteric coronaviruses, specific primers and probes were designed based on the highly conserved regions of transmissible gastroenteritis virus (TGEV) N, porcine epidemic diarrhea virus (PEDV) M, porcine deltacoronavirus (PDCoV) M, and porcine enteric alphacoronavirus (PEAV) N genes respectively. To develop a simple, rapid, accurate, and high-throughput detection method for diagnosis and differential diagnosis on swine enteric coronaviruses, a TaqMan-probe-based multiplex real-time RT-qPCR assay was established to simultaneously detect TGEV, PEDV, PEAV, and PDCoV from the same reaction vial. To establish a high-specific multiplex real-time RT-qPCR for the detection and differential diagnosis of swine enteric coronaviruses, primer sets, and TaqMan probes were designed targeting the highly conserved regions of PEDV or PDCoV M genes and TGEV or PEAV N genes, based on bioinformatics analysis of each virus.
cord-276575-jfug80yu	2007	
cord-282321-svoshzz8	2016	
cord-287602-vda01gj6	2018	
cord-288673-ku3tmjd3	2012	Because proteases play essential roles in life and death processes in all living organisms and because peptide bond hydrolysis is irreversible, anomalies in proteolytic activities lead to numerous pathological conditions, including cancer, neurodegenerative disorders and inflammatory and cardiovascular diseases, as well as bacterial, viral and parasitic diseases (Lopez-Otin and Bond 2008; Turk 2006) . Another important oral cavity pathogen involved in periodontal disease, Porphyromonas gingivalis, in addition to a few cysteine proteases (discussed further in the following), produces a serine protease, a prolyl tripeptidyl peptidase PtpA (family S9), which is involved in degrading host connective tissue, providing nutrients for bacterial growth (Banbula et al. Several proteases of the serine catalytic type have also been targeted for the design of specific protease inhibitors for use in cancer treatment, including the urokinase plasminogen activator and matriptase (Abbenante and Fairlie 2005; Bialas and Kafarski 2009; Ulisse et al.
cord-302503-7s9f8wje	2020	In October 2010, a severe PED outbreak caused by a highly virulent PEDV variant emerged in southern China with high mortality ranging from 70 to 100%; the result was devastating damage to the pig farm industry and tremendous economic losses, and later, the PEDV variant spreads to other countries, e.g., USA, Canada, and Mexico ( For the early and rapid detection of PEDV, different types of PCR methods have been developed. A TaqMan probe-based real-time PCR to differentiate porcine epidemic diarrhea virus virulent strains from attenuated vaccine strains Development and evaluation of a duplex real-time RT-PCR for detection and differentiation of virulent and variant strains of porcine epidemic diarrhea viruses from the United States Development of a TaqMan-based real-time RT-PCR assay for the detection of SADS-CoV associated with severe diarrhea disease in pigs
cord-321602-88b2h06y	2020	
cord-339694-sp212tai	2016	
cord-348310-nc1tq5af	2019	
cord-354904-7gq2e6f0	2018