Escherichia coli expressing F4 fimbriae is the major pathogenic bacteria that causes diarrhea in piglets before weaning. The adhesion of E. coli to the brush borders of the epithelial cells of piglets is the precondit...Escherichia coli expressing F4 fimbriae is the major pathogenic bacteria that causes diarrhea in piglets before weaning. The adhesion of E. coli to the brush borders of the epithelial cells of piglets is the precondition leading to diarrhea, which in turn is due to the presence of the F4 receptors determined by an autosomal recessive gene on the brush borders of the epithelial cells. In order to clarify the genetic mechanism of the adhesion, an in vitro adhesion experiment was carded out for three variants of E. coli F4 (ab, ac, and ad) in 366 piglets of three pig breeds [Landrace (LR), Large White (LW), and Songliao Black (SB)]. The results showed that there existed significant differences (P〈0.001) in the adhesion percentage among the three breeds. Most SB piglets were nonadhesive for all the three variants, whereas most LR piglets were adhesive. Within each breed except for LR, the proportions of the three F4 variants adhering to the brush borders differed significantly. According to the patterns of the adhesion of the three F4 variants in the three breeds, it is very likely that the three F4 variants F4ab, F4ac, and F4ad have different receptors that are controlled by three different loci.展开更多
The morphology and structure of the olfactory organ of Cynoglossus semilaevis Gunther are described. The oval olfactory sacs on both sides differ in size and in the number of lamellae, With those on the abocular side ...The morphology and structure of the olfactory organ of Cynoglossus semilaevis Gunther are described. The oval olfactory sacs on both sides differ in size and in the number of lamellae, With those on the abocular side having smaller sacs and fewer lamellae than those on the ocular side. On the ocular side, the average ratio of sac length to eye diameter is 2.1 (i.e.〉1) with an average of 91 lamellae, while on the abocular side, the values were 1.7 (i.e.〉1) and 69, respectively. In addition, the surface morphology varies in different parts of the lamella. The frontal part, near the anterior nostril, is a non-sensory margin with cilia-free epidermal cells. Within this is an internal ciliated sensory area, which is intercalated with ciliated receptor cells and a few ciliated non-sensory cells. Additionally, some dense ciliated non-sensory cells make up a non-sensory area, which also contains cilia-free epidermal cells distributed in patches. In the rear of the olfactory sac near the posterior nostril, the lamellae differ in morphology from those of the frontal olfactory sac but are similar in having few ciliated receptor cells. In other words, the surface of the lamellae in the rear part of the olfactory sac is mainly non-sensory. At present, four types of lamellae (~ E IlIand IV) have been recognized in relation to the pattern of the sensory epithelium. In this study, the frontal and rear lamellae resembled types I and IV, respectively, but are referred to as types r and IV because they are slightly less developed. Data on the ratio of length of lamellae to eye diameter, number of lamellae and the type of surface pattern of the lamellae show that the development of the olfactory system of C. semilaevis facilitates prey capture.展开更多
There are four types of gastric carcinoid tumors, classified according to their histology and malignant potential. Only a few cases of carcinoid tumors in patients infected with Helicobacter pylori (H. pylor/) have ...There are four types of gastric carcinoid tumors, classified according to their histology and malignant potential. Only a few cases of carcinoid tumors in patients infected with Helicobacter pylori (H. pylor/) have been reported so far. We report a patient infected with H. pylori presenting with a small solitary gastric carcinoid tumor with very low proliferative rate and normal gas- trin levels. The tumor was endoscopically removed and the patient received an eradication therapy against H. pylori. No signs of metastatic disease have been found so far during more than 3 year of follow-up. Infection with H. pylori may cause chronic gastritis with normal or elevated gastrin levels, leading to the develop- ment of gastric carcinoids by mechanisms unrelated to gastrin. Enterochromaffin-like cell tumors related to a chronic H. pylori infection may be considered as a distinct type of gastric carcinoid tumors.展开更多
AIM: To detect and evaluate the antibodies against Helicobacter pylori (H pylori) neutrophil-activating protein (HP-NAP) in patients with gastric cancer and other gastroduodenal diseases.METHODS: Recombinant HP-...AIM: To detect and evaluate the antibodies against Helicobacter pylori (H pylori) neutrophil-activating protein (HP-NAP) in patients with gastric cancer and other gastroduodenal diseases.METHODS: Recombinant HP-NAP was prepared from a prokaryotic expression system in Escherichia coll. Serum positivity and level of HP-NAP-specific antibodies in sera from 43 patients with gastric cancer, 28 with chronic gastritis, 28 with peptic ulcer, and 89 healthy controls were measured by rHP-NAP-based ELISA. rHP-NAP-stimulated production of interleukin-8 (IL-8) and growth-related oncogene (GROα) cytokines in the culture supernatant of SGC7901 gastric epithelial cells was also detected.RESULTS: The serum positivity and mean absorbancevalue of HP-NAP-specific antibodies in the gastriccancer group (97.7% and 1.01 ± 0.24) were significantly higher than those in the chronic gastritisgroup (85.7% and 0.89 ± 0.14, P 〈 0.005) and healthy control group (27.7% and 0.65 ± 0.18, P 〈 0.001). The sensitivity and specificity of ELISA for the detection of HP-NAP-specific antibodies were 95.5% and 91.5%, respectively. HP-NAP could slightly upregulate IL-8 production in gastric epithelial cell lines but had no effect on GROα production.CONCLUSION: Infection with virulent H py/ori strains secreting HP-NAP is associated with severe gastroduodenal diseases, and HP-NAP may play a role in the development of gastric carcinoma, rHP-NAP- based ELISA can be used as a new method to detect H pylori infection. The direct effect of HP-NAP on gastric epithelial cells may be limited, but HP-NAP may contribute to inflammatory response or carcinogenesis by activating neutrophils.展开更多
PROTEUS syndrome is characterized by patchy and progressive overgrowth affecting multiple tissues, including bone, soft tissue, and skin, along with susceptibility to the development oftumors. It was originally descri...PROTEUS syndrome is characterized by patchy and progressive overgrowth affecting multiple tissues, including bone, soft tissue, and skin, along with susceptibility to the development oftumors. It was originally described by Cohen and Hayden in 19791 and named by Wiedmann in 1983.2 The cause of Proteus syndrome is proposed to be a somatic mutation that is lethal in non-mosaic state; cells derived from the mutated cell line carrying this mutation result in anomalies in multiple tissues.3 The clinical manifestations are variable,展开更多
Cytochrome P450nor gene was cloned into the expression vector pET-28 to yield the recombinant expression plasmid pET-P450nor, which could direct the synthesis of a eukaryotic derived protein in Escherichia coli BL21. ...Cytochrome P450nor gene was cloned into the expression vector pET-28 to yield the recombinant expression plasmid pET-P450nor, which could direct the synthesis of a eukaryotic derived protein in Escherichia coli BL21. The vector allows overproduction and single-step purification of (His) 6-tagged cytochrome P450nor by the facilitation of metal (Ni 2+ ) chelate affinity chromatography. The expression level of cytochrome P450nor was high at 30℃ after IPTG induction. SDS-PAGE and Western blot analysis showed a specific band (about 43 kDa). The overproduced cytochrome P450nor was purified to electrophoretic homogeneity within 2.5 h and about 20.8 mg purified protein was obtained from 2 L cell culture. The proliferation of SSMC-7721 cell line could be inhibited by cytochrome P450nor. Rabbit polyclonal antibodies (titer over 64 000 ) were produced against recombinant cytochrome P450nor and proved to be very useful for immunoblotting study. Availability of this expression system and specific antibodies should facilitate characterization of the role of cytochrome P450nor in the metabolism of NO.展开更多
The mature Shiga toxin B (StxB) gene was optimized and generated by overlapping PCR. Recombinant expression vector pQE40-DHFR/StxB was constructed when the gene was cloned into pQE fusion expression vector. Induced ...The mature Shiga toxin B (StxB) gene was optimized and generated by overlapping PCR. Recombinant expression vector pQE40-DHFR/StxB was constructed when the gene was cloned into pQE fusion expression vector. Induced by Isopropyl β-D-thiogalactoside (IPTG), the DHFR/StxB fusion protein was highly expressed to the level of 41.36% in E. coli MI5 cells. The 35 kDa fusion protein with a 6 His-tag was one-step purified from inclusion bodies using Ni-NTA affinity chromatography column under denaturing conditions, and was refolded by dialyzing with a decreasing urea gradient. Purified DHFR/StxB fusion protein was used to immunize Kunming mice for generating the ascitic polyclonal antibody against recombinant StxB protein by injecting sarcoma 180 cells and the titer ofascitic polyclonal antibody is up to 1: 1× 10^6 detected by the indirect enzyme linked immunosorbent assy (ELISA). Western immunoblotting analysis revealed that the ascitic polyclonal antibody against StxB had a specific affinity for a 70 kDa shiga toxin protein of Shigella dysenteriae type 1. It is a new simple and quick method to produce a large amount of ascitic polyclonal antibody. The antibody is used to develop immunological method for detecting shiga toxin.展开更多
Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors known to play a pivotal role in regulations of metabolism. In order to yield soluble ligand binding domain of PPARδ (P...Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors known to play a pivotal role in regulations of metabolism. In order to yield soluble ligand binding domain of PPARδ (PPARδLBD) for screening ligands, the cDNA was amplified using total RNA from HepG2 cells by RT-PCR. Then the enzyme-digested product was inserted . downstream of the malE gene in the vectorpMAL-p2x, which encoded maltose-binding protein (MBP), resulting in the expression of an MBP-PPARδLBD fusion protein. The recombinant plasmid was transformed into E. coli TBI that was cultured shakily at 30 ℃, 200 r/min and induced by 0.4 mmol/L IPTG for 6 h. The cells were harvested by centrifugation and broken by sonication. The expressed fusion protein was soluble and accounted for 0.31 of the total protein in the supernatant. Western blot analysis showed that the expressed MBP-PPARδLBD could bind to anti-MBP-antibody. The MBP-PPARδLBD fusion protein of 77 kDa and the PPARδLBD protein of 34 kDa were obtained by amylose-resin affinity chromatography without or with digestion of Factor Xa. They were both homogeneity, judged by SDS-PAGE. The recombinant MBP-PPARδLBD and PPARδLBD protein with high purity is obtained, which provides the necessary material for screening and researching PPARδ ligands.展开更多
To obtain the P8 protein of Rice gall dwarf virus (RGDV) with biological activity,its outer coat protein gene S8 was expressed in Spodoptera frugiperda (Sf9) insect cells using the baculovirus expression system.The S8...To obtain the P8 protein of Rice gall dwarf virus (RGDV) with biological activity,its outer coat protein gene S8 was expressed in Spodoptera frugiperda (Sf9) insect cells using the baculovirus expression system.The S8 gene was subcloned into the pFastBacTM1 vector,to produce the recombinant baculovirus transfer vector pFB-S8.After transformation,pFB-S8 was introduced into the competent cells (E.coli DH10Bac) containing a shuttle vector,Bacmid,generating the recombinant bacmid rbpFB-S8.After being infected by recombinant baculovirus rvpFB-S8 at different multiplicities of infection,Sf9 cells were collected at different times and analyzed by SDS-PAGE,Western blotting and immunofluorescence microscopy.The expression level of the P8 protein was highest between 48-72 h after transfection of Sf9 cells.Immunofluorescence microscopy showed that P8 protein of RGDV formed punctate structures in the cytoplasm of Sf9 cells.展开更多
The multiple antibiotic resistance regulatory protein(MarR) binds to two promoter sites on the marO operator in Escherichia coli.Our study showed that more than one MarR dimer proteins bound to either of its two promo...The multiple antibiotic resistance regulatory protein(MarR) binds to two promoter sites on the marO operator in Escherichia coli.Our study showed that more than one MarR dimer proteins bound to either of its two promoter sites(Site I and Site II),suggesting that MarR might form higher complexes than homodimers when bound to DNA inside E.coli cells.To further verify this hypothesis,we site-specifically incorporated a photocrosslinking probe at the interface between two MarR dimer proteins.Photolysis in living E.coli cells revealed a covalent linkage between the two interdimer subunits of MarR,suggesting that MarR forms dimer of dimers in vivo.展开更多
Fibroblast growth factor(FGF) is found throughout multicellular organisms; however, fgf homologs(vfgf) have only been identified among viruses in lepidopteran baculoviruses. The function of v FGFs from Group I alphaba...Fibroblast growth factor(FGF) is found throughout multicellular organisms; however, fgf homologs(vfgf) have only been identified among viruses in lepidopteran baculoviruses. The function of v FGFs from Group I alphabaculoviruses, including Autographa californica multiple nucleopolyhedrovirus(Ac MNPV) and Bombyx mori nucleopolyhedrovirus(Bm NPV), involves accelerated killing of infected larvae by both viruses. The v FGF of Group II alphabaculovirus is structurally different from that of Group I alphabaculovirus, with a larger C-terminal region and additional N-linked glycosylation sites. In this study, we characterized the Group II alphabaculovirus v FGF of Helicoverpa armigera single nucleopolyhedrovirus(Hear NPV). The transcription and expression of vfgf was detected at 3 h and 16 h post-infection in Hear NPV-infected cells. To further study v FGF function, we constructed vfgf-knockout and-repaired Hear NPV bacmids and investigated their affect in both cultured cells and insects. Deletion of vfgf had no effect on budded-virus production or viral DNA replication in cultured Hz AM1 cells. However, bioassays showed that Hear NPV vfgf deletion significantly increased the median lethal dose and delayed the median lethal time by ~12 h in the host insect when the virus was delivered orally. These results suggested that v FGF is an important virulent factor for HearN PV infection and propagation in vivo.展开更多
基金National Basic Research Program of China (No. 2006CB102104)National Natural Sci-ences Foundation of China (No. 30430500).
文摘Escherichia coli expressing F4 fimbriae is the major pathogenic bacteria that causes diarrhea in piglets before weaning. The adhesion of E. coli to the brush borders of the epithelial cells of piglets is the precondition leading to diarrhea, which in turn is due to the presence of the F4 receptors determined by an autosomal recessive gene on the brush borders of the epithelial cells. In order to clarify the genetic mechanism of the adhesion, an in vitro adhesion experiment was carded out for three variants of E. coli F4 (ab, ac, and ad) in 366 piglets of three pig breeds [Landrace (LR), Large White (LW), and Songliao Black (SB)]. The results showed that there existed significant differences (P〈0.001) in the adhesion percentage among the three breeds. Most SB piglets were nonadhesive for all the three variants, whereas most LR piglets were adhesive. Within each breed except for LR, the proportions of the three F4 variants adhering to the brush borders differed significantly. According to the patterns of the adhesion of the three F4 variants in the three breeds, it is very likely that the three F4 variants F4ab, F4ac, and F4ad have different receptors that are controlled by three different loci.
基金Supported by Qingdao Natural Science Foundation (No. 09-1-3-10-jch)"Taishan Scholar" Project Special Fundthe Yellow Sea Fisheries Research Institute Scientific and Research Fund
文摘The morphology and structure of the olfactory organ of Cynoglossus semilaevis Gunther are described. The oval olfactory sacs on both sides differ in size and in the number of lamellae, With those on the abocular side having smaller sacs and fewer lamellae than those on the ocular side. On the ocular side, the average ratio of sac length to eye diameter is 2.1 (i.e.〉1) with an average of 91 lamellae, while on the abocular side, the values were 1.7 (i.e.〉1) and 69, respectively. In addition, the surface morphology varies in different parts of the lamella. The frontal part, near the anterior nostril, is a non-sensory margin with cilia-free epidermal cells. Within this is an internal ciliated sensory area, which is intercalated with ciliated receptor cells and a few ciliated non-sensory cells. Additionally, some dense ciliated non-sensory cells make up a non-sensory area, which also contains cilia-free epidermal cells distributed in patches. In the rear of the olfactory sac near the posterior nostril, the lamellae differ in morphology from those of the frontal olfactory sac but are similar in having few ciliated receptor cells. In other words, the surface of the lamellae in the rear part of the olfactory sac is mainly non-sensory. At present, four types of lamellae (~ E IlIand IV) have been recognized in relation to the pattern of the sensory epithelium. In this study, the frontal and rear lamellae resembled types I and IV, respectively, but are referred to as types r and IV because they are slightly less developed. Data on the ratio of length of lamellae to eye diameter, number of lamellae and the type of surface pattern of the lamellae show that the development of the olfactory system of C. semilaevis facilitates prey capture.
文摘There are four types of gastric carcinoid tumors, classified according to their histology and malignant potential. Only a few cases of carcinoid tumors in patients infected with Helicobacter pylori (H. pylor/) have been reported so far. We report a patient infected with H. pylori presenting with a small solitary gastric carcinoid tumor with very low proliferative rate and normal gas- trin levels. The tumor was endoscopically removed and the patient received an eradication therapy against H. pylori. No signs of metastatic disease have been found so far during more than 3 year of follow-up. Infection with H. pylori may cause chronic gastritis with normal or elevated gastrin levels, leading to the develop- ment of gastric carcinoids by mechanisms unrelated to gastrin. Enterochromaffin-like cell tumors related to a chronic H. pylori infection may be considered as a distinct type of gastric carcinoid tumors.
基金Supported by Grants from Guangdong Natural Science Foundation Project,5004750National Key Development Project,973 Program 2002CB513206
文摘AIM: To detect and evaluate the antibodies against Helicobacter pylori (H pylori) neutrophil-activating protein (HP-NAP) in patients with gastric cancer and other gastroduodenal diseases.METHODS: Recombinant HP-NAP was prepared from a prokaryotic expression system in Escherichia coll. Serum positivity and level of HP-NAP-specific antibodies in sera from 43 patients with gastric cancer, 28 with chronic gastritis, 28 with peptic ulcer, and 89 healthy controls were measured by rHP-NAP-based ELISA. rHP-NAP-stimulated production of interleukin-8 (IL-8) and growth-related oncogene (GROα) cytokines in the culture supernatant of SGC7901 gastric epithelial cells was also detected.RESULTS: The serum positivity and mean absorbancevalue of HP-NAP-specific antibodies in the gastriccancer group (97.7% and 1.01 ± 0.24) were significantly higher than those in the chronic gastritisgroup (85.7% and 0.89 ± 0.14, P 〈 0.005) and healthy control group (27.7% and 0.65 ± 0.18, P 〈 0.001). The sensitivity and specificity of ELISA for the detection of HP-NAP-specific antibodies were 95.5% and 91.5%, respectively. HP-NAP could slightly upregulate IL-8 production in gastric epithelial cell lines but had no effect on GROα production.CONCLUSION: Infection with virulent H py/ori strains secreting HP-NAP is associated with severe gastroduodenal diseases, and HP-NAP may play a role in the development of gastric carcinoma, rHP-NAP- based ELISA can be used as a new method to detect H pylori infection. The direct effect of HP-NAP on gastric epithelial cells may be limited, but HP-NAP may contribute to inflammatory response or carcinogenesis by activating neutrophils.
文摘PROTEUS syndrome is characterized by patchy and progressive overgrowth affecting multiple tissues, including bone, soft tissue, and skin, along with susceptibility to the development oftumors. It was originally described by Cohen and Hayden in 19791 and named by Wiedmann in 1983.2 The cause of Proteus syndrome is proposed to be a somatic mutation that is lethal in non-mosaic state; cells derived from the mutated cell line carrying this mutation result in anomalies in multiple tissues.3 The clinical manifestations are variable,
基金This work was supported by National Natural Science Foundation of Chi na (No. 30170011)
文摘Cytochrome P450nor gene was cloned into the expression vector pET-28 to yield the recombinant expression plasmid pET-P450nor, which could direct the synthesis of a eukaryotic derived protein in Escherichia coli BL21. The vector allows overproduction and single-step purification of (His) 6-tagged cytochrome P450nor by the facilitation of metal (Ni 2+ ) chelate affinity chromatography. The expression level of cytochrome P450nor was high at 30℃ after IPTG induction. SDS-PAGE and Western blot analysis showed a specific band (about 43 kDa). The overproduced cytochrome P450nor was purified to electrophoretic homogeneity within 2.5 h and about 20.8 mg purified protein was obtained from 2 L cell culture. The proliferation of SSMC-7721 cell line could be inhibited by cytochrome P450nor. Rabbit polyclonal antibodies (titer over 64 000 ) were produced against recombinant cytochrome P450nor and proved to be very useful for immunoblotting study. Availability of this expression system and specific antibodies should facilitate characterization of the role of cytochrome P450nor in the metabolism of NO.
文摘The mature Shiga toxin B (StxB) gene was optimized and generated by overlapping PCR. Recombinant expression vector pQE40-DHFR/StxB was constructed when the gene was cloned into pQE fusion expression vector. Induced by Isopropyl β-D-thiogalactoside (IPTG), the DHFR/StxB fusion protein was highly expressed to the level of 41.36% in E. coli MI5 cells. The 35 kDa fusion protein with a 6 His-tag was one-step purified from inclusion bodies using Ni-NTA affinity chromatography column under denaturing conditions, and was refolded by dialyzing with a decreasing urea gradient. Purified DHFR/StxB fusion protein was used to immunize Kunming mice for generating the ascitic polyclonal antibody against recombinant StxB protein by injecting sarcoma 180 cells and the titer ofascitic polyclonal antibody is up to 1: 1× 10^6 detected by the indirect enzyme linked immunosorbent assy (ELISA). Western immunoblotting analysis revealed that the ascitic polyclonal antibody against StxB had a specific affinity for a 70 kDa shiga toxin protein of Shigella dysenteriae type 1. It is a new simple and quick method to produce a large amount of ascitic polyclonal antibody. The antibody is used to develop immunological method for detecting shiga toxin.
基金Supported by the National Natural Science Foundation of China (30572353)
文摘Peroxisome proliferator-activated receptors (PPARs) are ligand-activated transcription factors known to play a pivotal role in regulations of metabolism. In order to yield soluble ligand binding domain of PPARδ (PPARδLBD) for screening ligands, the cDNA was amplified using total RNA from HepG2 cells by RT-PCR. Then the enzyme-digested product was inserted . downstream of the malE gene in the vectorpMAL-p2x, which encoded maltose-binding protein (MBP), resulting in the expression of an MBP-PPARδLBD fusion protein. The recombinant plasmid was transformed into E. coli TBI that was cultured shakily at 30 ℃, 200 r/min and induced by 0.4 mmol/L IPTG for 6 h. The cells were harvested by centrifugation and broken by sonication. The expressed fusion protein was soluble and accounted for 0.31 of the total protein in the supernatant. Western blot analysis showed that the expressed MBP-PPARδLBD could bind to anti-MBP-antibody. The MBP-PPARδLBD fusion protein of 77 kDa and the PPARδLBD protein of 34 kDa were obtained by amylose-resin affinity chromatography without or with digestion of Factor Xa. They were both homogeneity, judged by SDS-PAGE. The recombinant MBP-PPARδLBD and PPARδLBD protein with high purity is obtained, which provides the necessary material for screening and researching PPARδ ligands.
基金supported by the National Science Foundation of China (30970135)The Key Project of Genetically Modified Organisms Breeding(2009ZX08009-044B)+1 种基金the Natural Science Foundation of Fujian Province of China (No.2006J0065)the Public-interest Scientific Institution Basal Research Fund of Fujian Province (2009R10029-3)
文摘To obtain the P8 protein of Rice gall dwarf virus (RGDV) with biological activity,its outer coat protein gene S8 was expressed in Spodoptera frugiperda (Sf9) insect cells using the baculovirus expression system.The S8 gene was subcloned into the pFastBacTM1 vector,to produce the recombinant baculovirus transfer vector pFB-S8.After transformation,pFB-S8 was introduced into the competent cells (E.coli DH10Bac) containing a shuttle vector,Bacmid,generating the recombinant bacmid rbpFB-S8.After being infected by recombinant baculovirus rvpFB-S8 at different multiplicities of infection,Sf9 cells were collected at different times and analyzed by SDS-PAGE,Western blotting and immunofluorescence microscopy.The expression level of the P8 protein was highest between 48-72 h after transfection of Sf9 cells.Immunofluorescence microscopy showed that P8 protein of RGDV formed punctate structures in the cytoplasm of Sf9 cells.
基金supported by research grants from the National Natural Science Foundation of China(91013005, 21001010 and 20932006 to P.R.C.)National Key Basic Research Foundation of China(2010CB912300 to P.R.C.)
文摘The multiple antibiotic resistance regulatory protein(MarR) binds to two promoter sites on the marO operator in Escherichia coli.Our study showed that more than one MarR dimer proteins bound to either of its two promoter sites(Site I and Site II),suggesting that MarR might form higher complexes than homodimers when bound to DNA inside E.coli cells.To further verify this hypothesis,we site-specifically incorporated a photocrosslinking probe at the interface between two MarR dimer proteins.Photolysis in living E.coli cells revealed a covalent linkage between the two interdimer subunits of MarR,suggesting that MarR forms dimer of dimers in vivo.
基金supported by grants from the National Science Foundation of China (No.31200124 and 31321001)the Strategic Priority Research Program of the Chinese Academy of Sciences (Grant No.XDB11030400)
文摘Fibroblast growth factor(FGF) is found throughout multicellular organisms; however, fgf homologs(vfgf) have only been identified among viruses in lepidopteran baculoviruses. The function of v FGFs from Group I alphabaculoviruses, including Autographa californica multiple nucleopolyhedrovirus(Ac MNPV) and Bombyx mori nucleopolyhedrovirus(Bm NPV), involves accelerated killing of infected larvae by both viruses. The v FGF of Group II alphabaculovirus is structurally different from that of Group I alphabaculovirus, with a larger C-terminal region and additional N-linked glycosylation sites. In this study, we characterized the Group II alphabaculovirus v FGF of Helicoverpa armigera single nucleopolyhedrovirus(Hear NPV). The transcription and expression of vfgf was detected at 3 h and 16 h post-infection in Hear NPV-infected cells. To further study v FGF function, we constructed vfgf-knockout and-repaired Hear NPV bacmids and investigated their affect in both cultured cells and insects. Deletion of vfgf had no effect on budded-virus production or viral DNA replication in cultured Hz AM1 cells. However, bioassays showed that Hear NPV vfgf deletion significantly increased the median lethal dose and delayed the median lethal time by ~12 h in the host insect when the virus was delivered orally. These results suggested that v FGF is an important virulent factor for HearN PV infection and propagation in vivo.