cloning expression and purification of rvfv n and nss

... 14kD NSs N Figure Expression and purification of the RVFV N and NSs antigens Expression and purification of the RVFV N and NSs antigens Protein samples were mixed with reducing sample buffer and ... countries where RVFV is prevalent Results and Discussion Cloning, expression and purification of RVFV N and NSs ELISAs have been used in the past in the diagnosis of RVFV infection in both humans ... and 1C) The induction of N was very tightly controlled, but as indicated by lane of Figure 1B, there was some leaky expression of NSs prior to induction Titration of antigens The N and NSs antigens...

Ngày tải lên: 12/08/2014, 04:21

... properties of Conus PDI have not been thoroughly investigated, especially using the endogenous conotoxins as substrates In this article, we report gene cloning, recombinant expression and enzymatic ... Biochemical and gene expression analyses of conotoxins in Conus textile venom ducts Biochem Biophys Res Commun 328, 362–367 12 Nielsen H, Engelbrecht J, Brunak S & von Heijne G (1997) Identification of ... amidation of the mature form of x-conotoxin containing an amidated C-terminus, as isolated from the venom of Conus magnus (x-MVIIA) can significantly increase the folding yield [6] The carboxylation of...

Ngày tải lên: 07/03/2014, 05:20

... Supernatants of isolates from patients infected with S pyogenes were analyzed by SDS/ PAGE and immunoblotting using a rabbit anti-SSA serum Lanes and 3: supernatants of isolates; Lanes and 4: supernatants ... contrary, no binding of Vb2.1 TCR to SSA was detected even using a 160 lM concentration of b chain Trials using higher concentrations were unsuccessful due to nonspecific aggregation of the Vb2.1 ... useful in the development of vaccines and in cancer therapy SAgs with mutated TCR and/ or MHC binding sites could be employed as vaccines against TSS and food poisoning to generate protective antibodies...

Ngày tải lên: 07/03/2014, 16:20

... cis-prenyltransferases, was termed LT600 3¢- and 5¢-RACE reaction and cloning of Hevea cis-prenyltransferase cDNA Cloning of the full-length cDNA of Hevea cis-prenyltransferase, used poly(A)+ RNA ... fragments and obtained two distinct cDNAs that contain sequences identical to those obtained by 3¢-RACE General cDNAs containing 5¢- and 3¢-untranslated regions were also amplified by RT-PCR and ... amino acid sequences of HRT1 (A) and HRT2 (B) Numbers of nucleotide sequence and amino acid sequence are indicated on the left and right, respectively The underline in HRT1 indicates the sequence...

Ngày tải lên: 07/03/2014, 21:20

... of the restriction endonuclease in the oligonucleotides are underlined Initiation and termination codons for translation are in bold Sequence Primer ZF SULT1 ST1 Sense Antisense 5¢-CGCGGATCCATGGACATGCCTGACTTTTCT-3¢ ... (Amount of enzyme and reaction time were chosen to ensure that there was no more than 5% reaction: the reaction was linear with time and amount of enzyme.) The reaction was terminated by heating ... Two Ôsignature sequencesÕ located in the N- terminal and C-terminal regions, and a conserved sequence in the middle region, are underlined They were then isolated and subjected to complete nucleotide...

Ngày tải lên: 08/03/2014, 02:20

... final concentrations: ampicillin (Amp), 100 lgÆmL)1; kanamycin (Km), 50 lgÆmL)1 DNA manipulation and transformation The isolation and manipulation of recombinant DNA were performed using standard ... Glu and Asp, and the formation of Glu and Asp is used to synthesize Gln and Asn, respectively These four residues are common components in lipopeptides, such as surfactin, iturin and fengycin Another ... Expression and purification of AATB3 and its mutants To produce recombinant AATB3 and the three mutant proteins, the aatB3 gene and its mutants were expressed in E coli The recombinant proteins...

Ngày tải lên: 14/03/2014, 23:20

... component of the M tuberculosis cell wall, and might also function as a stabilizer and protector of membranes and proteins when this organism undergoes latency Therefore, enzymes involved in the ... TPS, and another sequence of about 16 amino acids starting at position 421 of the M tuberculosis TPS Isolation and purification of recombinant TPS To maximize conditions for production of recombinant ... by centrifugation, suspended in 50 mM NaH2PO4 pH 8.0, containing 300 mM NaCl and 10 mM imidazole, and disrupted by sonication The cell debris were removed by centrifugation and the supernatant...

Ngày tải lên: 17/03/2014, 10:20

... His-binding resin was from Novagen Except where otherwise specified, all DNA manipulation enzymes, including restriction endonucleases, polymerases and ligase, were from New England Biolabs and were ... amount of the product, trehalose, increased with increasing concentrations of maltose in the incubation up to about mM, and then leveled off with further increases in substrate concentration When ... proportional to the amount of enzyme added to the incubation mixtures (data not shown) The formation of trehalose from maltose was also linear with time of incubation and enzyme concentration, but...

Ngày tải lên: 23/03/2014, 13:20

... sequence and it has a convenient NotI site at its C terminus, which allows cloning in-frame with alanine as the first amino acid of the mature protein An alignment of the amino acid sequences of ... Irwin, D., Barr, B.K., Benjamin, A & Wilson, D.B (1993) DNA sequences and expression in Streptomyces lividans of an exoglucanase gene and an endoglucanase gene from Thermomonospora fusca Appl Environ ... fragments of pET26B+ (Novagen) digested with NdeI and XhoI, a NdeI–NotI fragment containing the T fusca Cel6A signal peptide and the above PCR fragment digested with NotI and XhoI This ligation mixture...

Ngày tải lên: 23/03/2014, 21:20

... ND ND ND ND 22.2 > 29.6 1.4 ND 5.6 11.1 5.6 ND ND 11.1 5.6 2.8 11.1 ND ND ND ND 22.2 > 29.6 1.4 ND > 22.3 ND ND ND ND ND ND ND > 22.3 ND 4.2 ND ND ND ND ND ND ND ND > 269.25 25.24 201.94 ND ND ... supernatant with IPTG; lane 0, protein standards (C) Protein bands after affinity chromatography and renaturing process Lanes and 2, protein bands after separation by affinity column; lanes and 4, ... 5¢-AAGCAGTGGTATCAACGCAGAGT-3¢ and CDS III ⁄ 3¢ PCR primer, using Advantage DNA Polymerase from Clontech Screening of cathelicidin-encoding cDNAs and phylogenetic tree construction On the basis of the conserved signal...

Ngày tải lên: 28/03/2014, 23:20

... gene product may be involved in the degradation of nitroalkanes and this gene was designated naoA (nitroalkane-oxidizing enzyme gene) Expression of naoA in E coli To study the function of the naoA ... Restriction enzymes and T4 DNA ligase were purchased from Boehringer-Mannheim and Sino-American Biotechnology Company (Luoyang, Chi1 na) DIG labeling and detection kits (Boehringer-Mannheim) were ... the common products of nitroalkane oxidation catalyzed by 2-nitropropane dioxygenase and nitroalkane oxidase, were detected in the oxidation of 1-nitropropane, 2-nitropropane and nitroethane catalyzed...

Ngày tải lên: 31/03/2014, 08:20

... (PBS containing 90% glycerol), and analyzed with Confocal laser scanning microscopy (CLSM, Nikon, Japan) The expression of gE protein in DPV-infected cells RNA expression of DPV gE in DPV-infected ... were individually transformed with the positive recombinant plasmid and used for protein expression Expression and Purification of the recombinant protein Expression of this fusion protein was ... cells using Western blotting, and analyzed the DPV gE gene transcription in DPV-infected cells using the real time PCR and RT-PCR Results Cloning of DPV gE gene and the correct recombinant plasmid...

Ngày tải lên: 12/08/2014, 04:20

... life And thank my parents for their endless caring, encouragement and understanding i Table of contents Table of contents Content Page ACKNOWLEDGEMENT i LIST OF FIGURES ii LIST OF TABLES v LIST OF ... H pylori and epidemiology of H pylori infection 2.3.4 Evolutionary change of DNA sequences 2.3.4.1 Synonymous and non-synonymous substitutions Synonymous substitution is the change of nucleotide ... different families of signal sequences (s1a, s1b, s1c and s2) and two different families of middle-region alleles (m1 and m2) (Van Doorn et al., 1999) Different combinations between s and m regions...

Ngày tải lên: 12/09/2015, 10:19

... Understanding of the nature of the interaction between Osh7p and Vps4p and future directions 108 4.6 Understanding the nature of Osh7p modification 109 4.7 Future directions of the project 110 CONCLUSION ... Zhang Qian, Shaochong and Li Phing for providing useful comments and technical assistance I would also like to thank Dr Alan Munn, Jihui, Sebastain and Vicky from Yeast Laboratory in Institute of ... protein-protein interactions and are generally found in cytoskeleton proteins and transcription factors Thus the structure of Osh1p and Osh2p is suggestive of being able to bind both a phosphoinositide...

Ngày tải lên: 03/10/2015, 20:57

... Molecular cloning and sequence of x-5 gliadin gene Many kinds of genes encoding gliadins such as a-gliadin, c-gliadin and x-gliadin, have been cloned from common wheat (T aestivum) and sequenced The ... with sera of the three patients The binding of IgE to nOG5 was completely inhibited by increasing amounts of rOG5C in all patients At an inhibitor concentration of 0.1 and lgÆmL)1, nOG5 inhibited ... important and the use of standardized recombinant proteins might reduce inaccurate diagnosis Some recombinant food allergens have been produced and the advantages of recombinant proteins have been...

Ngày tải lên: 20/02/2014, 01:20

... Cloning and sequencing of caspase in rainbow trout, Oncorhynchus mykiss, and analysis of its expression under conditions known to induce apoptosis Dev Comp Immunol 25, 303–312 25 Mosmann, T (1994) ... production in mice, chimpanzees, baboons and humans [32–35] The presence of endogenous IL-10 confers protection from the lethal effects of endotoxin challenge and reduces the levels of TNF, IFN-c and ... expression and its role in the regulation of other pro-inflammatory genes, such as TNF and IFN, in fish, need to be conducted In conclusion, the IL-10 gene from carp has been isolated and its genomic...

Ngày tải lên: 20/02/2014, 02:21

... ecdysozoan are necessary for precise understanding of their physiology In conclusion, we successfully performed cDNA cloning of EcR and USP of L decemlineata, and in vitro translation of corresponding ... analysis of mRNA expression profile of L decemlineata, EcR and USP, and (c) the measurement of the binding affinity of steroidal and nonsteroidal ecdysone agonists to the in vitro translated receptor ... receptor-binding 4121 Molting hormone receptors of L decemlineata T Ogura et al Fig Comparison of the ligand binding domain sequence of EcRs Amino-acid sequences are compared for ligand binding domain of...

Ngày tải lên: 07/03/2014, 21:20

... re¯ect geranylgeranylation of the proteins For unknown reasons, HEL cells did not contain detectable amounts of Rab31 and Rab32, using immunoblotting techniques Rab31 protein was found in both MEG-01 ... positions of prestained protein standards are shown on the right membrane fractions and that no Rab31 or Rab32, and only a small amount of Rab11A, was present in the supernatant (cytosol) fraction ... proteins Thus, human and mouse Rab11A are 100% identical and human and mouse Rab5A 97% identical The main sequence differences between human Rab32 and the related mouse protein are in the N- and...

Ngày tải lên: 08/03/2014, 16:20

... comparison with that of an identical enzyme incubation mixture containing no inhibitor Results Cloning and sequence analysis of full-length tryptase cDNAs A partial cDNA (690 bp) encoding a new bovine ... Results) Organization of bovine tryptase genes and location of intron II–V Intron II–V length of the two genes encoding bovine tryptases was evaluated by amplification of bovine genomic DNA, using the ... tryptase isoform (BLT) was obtained from lung mRNA by RT-PCR, using primers N9 and N1 0, and by subsequent cloning and sequencing Based on this partial sequence, 5¢ RACE experiments and RT-PCR (using...

Ngày tải lên: 17/03/2014, 09:20

... b-actin nucleotide sequence Target sequence Sense and antisense oligonucleotide primers SULT1 ST1 Sense: Antisense: Sense: Antisense: Sense: Antisense: Sense: Antisense: Sense: Antisense: Sense: Antisense: ... compounds Specific activity (nmolÆmin)1Æmg)1) 3,3¢,5-Triiodo-D-Thyronine (D-T3) 24.32 17.74 17.39 11.62 4.31 NDb ND ND ND ND ND ND ND ND ND ND ND ND ND ND Chlorogenic acid Kaempferol Gallic acid Genistein ... and subcloned into pSTBlue-1 cloning vector and subjected to nucleotide sequencing [25] The nucleotide and deduced amino acid sequences of the cDNA were analyzed using blast search for sequence...

Ngày tải lên: 23/03/2014, 15:20