extracellular production isolation and molecular characterization

... puri, purified proteins; M, molecular weight standards For the molecular weight standards, the bands from top to bottom correspond to 116.0, 66.2, 45.0, 35.0, 25.0 and 18.4 kDa, respectively consistent ... extracts and the supernatant and precipitate fractions are shown in Fig It was found that E coli BL21(DE3) ⁄ pHYDJs and E coli BL21(DE3) ⁄ pHYDPf produced a predominant band with an apparent molecular ... standard molecular weight proteins The flow rate was set to 0.8 mLÆmin)1 The subunit molecular mass was also estimated by SDS–PAGE Characterization and comparative analyses of HYDJs and HYDBp...

Ngày tải lên: 18/02/2014, 08:20

... isolation 3.4.4 DNA sequencing and analysis 41 41 41 43 44 3.5 Probe labeling 3.5.1 DNA probes 3.5.2 RNA probes 44 44 46 3.6 Genomic DNA isolation and Southern analysis 46 3.7 RNA isolation and ... expressing sense, antisense and double-stranded GST cDNAs 5.2.6 Flowering and stress response in transgenic plants 5.2.7 Shoot regeneration response and ethylene production of cultured tissues ... antisense (GSTA4) and double-stranded (GST-DS1) cDNAs 112 Figure 23 Phenotype of WT and different transgenic Arabidopsis plants expressing sense (GST-S6), antisense (GST-A4) and doublestranded (GST-DS1)...

Ngày tải lên: 17/09/2015, 17:19

... C6 and C9 aldehydes (and the corresponding alcohol) were detected in all the samples and, similarly to the pattern of LOX expression and activity, their levels were higher in stage I seed and ... the shelf life and the organoleptic features of hazelnut seeds, we carried out a study to characterize hazelnut lipoxygenases at the biochemical and molecular level Materials and methods Plant ... NaCl/Cit 65 °C and chemiluminescent detection was carried out using CDP-STAR as substrate according to the manufacturer’s protocol (Roche) Results Lipoxygenase gene isolation and characterization...

Ngày tải lên: 21/02/2014, 00:20

... Corresponding pH optima and specific activities for syringaldazine and 2,6-DMP were pH 5.6 and 3.4 UÆmg protein)1 and pH 4.6 and 2.8 UÆmg protein)1, respectively In standard assay mixtures, the ... unaffected [24] HBT, XYL, FA and VA all induced extracellular laccase production in P sajor-caju and transcription levels of three laccase genes were increased by FA and XYL [5] Higher levels of ... agarose/Tris borate/EDTA gels and appropriate bands excised and purified from the gel using the NucleoTrap Gel Extraction Kit (Clontech) The purified DNA was precipitated with ethanol and resuspended in 10...

Ngày tải lên: 07/03/2014, 15:20

... described in the Materials and methods phenolic compounds and alginates, etc and hence preserved the enzymatic activity of lamjapin But the efficiency of extraction was still low and the proteins could ... Fast and sensitive detection of protein and DNA bands by treatment with potassium permanganate J Biochem Biophys Methods 11, 13–20 Neuhoff, V., Stamm R & Eibl, H (1985) Clear background and highly ... proteins and also to the low efficiency of extraction Physical properties of lamjapin Lamjapin obtained from the Mono Q column migrated as a single band characterized by both 12% SDS/PAGE and isoelectric...

Ngày tải lên: 08/03/2014, 10:20

... Trichoderma: isolation and expression of a full-length and a truncated mutant form Mol Gen Genet 251, 451–460 52 Tempelaars, C.A., Birch, P.R., Sims, P.F & Broda, P (1994) Isolation, characterization, and ... ATTCCGACTAC-3¢; and inner 3¢-RACE primer 5¢-AAC TCCTACGTGACCTACTCGAAC-3¢ PCR products were cloned and sequenced as described previously Fungal strain and growth conditions Isolation of cel7 cDNA and genomic ... which are stacked face-to-face and are highly curved, forming convex and concave surfaces The convex and concave sheets of the b-sandwich are composed of seven b-strands Many of the side-chains...

Ngày tải lên: 23/03/2014, 13:20

... exists as a monomer and dimer and splits into a membrane and soluble subcomplexes Separation of the Ndh complex by CN / PAGE (1D) and Tricine / PAGE (2D) and CN / PAGE (1D) and BN / PAGE (2D) ... The MS (left) and BS (right) chloroplasts were separated in CN ⁄ PAGE (1D, the horizontal bands) and then in nondenaturing conditions by BN ⁄ PAGE (2D) The molecular mass standards and direction ... (2D) The molecular mass standards and direction of migration are indicated for both the first (1D) and second (2D) dimension (A) Tricine ⁄ PAGE gels that resulted in 2D were electroblotted and immunodecorated...

Ngày tải lên: 23/03/2014, 13:20

... Lys56; and aD, Ala90–Lys108); (e) two short strands 4550 of an antiparallel b-sheet (b-wing) (b2, Ser74–Ser78; and b3, Glu81–Asn85); and (f) a C-terminal loop containing a short b4-strand (His115–Asn117) ... elements labels are indicated a-Helices and b-strands are colored in red and yellow, respectively The catalytic network and the Ca2+-binding residues are indicated and shown as sticks (B) The Ca2+-binding ... Effect of temperature and pH on ChPLA2 activity (A,C) and stability (B,D) ChPLA2 ( ) and PPLA2 ( ), used as controls, were tested for activity at various temperatures (A) and pH values (C), as...

Ngày tải lên: 30/03/2014, 01:20

... estimate their molecular sizes By calibration of the column with standard proteins of known molecular masses, the molecular masses of native FerA and FerB were estimated as 25 kDa and Table Purification ... values of Km and kcat are: (0.4 ± 0.2) mM and (0.91 ± 0.02) s)1 for Fe(III)EGTA (1.6 ± 0.8) mM and (0.16 ± 0.03) s)1 for Fe(III)EDTA (0.4 ± 0.2) mM and (0.08 ± 0.01) s)1 for Fe(III)citrate, and (0.4 ... chromophore uniquely attributable to this enzyme On the other hand, the solution of FerB was yellowish in color and exhibited absorption bands at 380 and 448 nm (Fig 3), diagnostic for the presence of a...

Ngày tải lên: 30/03/2014, 13:20

... and digested with EcoRV for 1.5 hrs at 37°C Unique bands generated by this restriction enzyme facilitate identification of single infections with EACMV and ACMV (lanes 1–3 and respectively) and ... analysis, cloning and nucleotide sequencing The studies described here demonstrate that FTA offers a simple, sensitive and specific tool appropriate for the diagnosis and molecular characterization ... sampling and recovery of viral pathogens from infected leaf tissues and their subsequent molecular analysis Utilising the geminiviruses that infect maize (Zea mays), cassava (Manihot esculenta) and...

Ngày tải lên: 19/06/2014, 08:20

... and digested with EcoRV for 1.5 hrs at 37°C Unique bands generated by this restriction enzyme facilitate identification of single infections with EACMV and ACMV (lanes 1–3 and respectively) and ... analysis, cloning and nucleotide sequencing The studies described here demonstrate that FTA offers a simple, sensitive and specific tool appropriate for the diagnosis and molecular characterization ... sampling and recovery of viral pathogens from infected leaf tissues and their subsequent molecular analysis Utilising the geminiviruses that infect maize (Zea mays), cassava (Manihot esculenta) and...

Ngày tải lên: 20/06/2014, 04:20

... (AF338717), Detection and molecular characterization of infectious bronchitis virus 221 GA/8077/99 (AF338718), DLD (EU589323), THA001 (DQ449628) Results Virus screening and isolation For virus ... Kumar S, Tamura K, Nei M MEGA3: Integrated software for Molecular Evolutionary Genetics Analysis and sequence Detection and molecular characterization of infectious bronchitis virus 223 alignment ... region of S1 gene covering HVR I and HVR II were amplified and used for typing the field isolates in Thailand Although many different strains of live attenuated and inactivated vaccines have been...

Ngày tải lên: 07/08/2014, 23:22

... CTC-3’ and 5’AAT GGA TTC TTG AAT GCT TCT GAA-3’ The resulting fragment was digested with Hind III and then ligated with PBI101.2, which was predigested with Hand III and Sma I For RAP2.4 and RAP2.4L, ... Growth of AtERF13, RAP2.4 and RAP2.4L knockout and RNAi lines RNAi lines of AtERF13 and knockout lines of RAP2.4 and RAP2.4L were prepared as described in the Methods, and their growth phenotypes ... equipments and plant growth facilities Authors’ contributions SL conducted the expression analysis and analyzed the OX and KO lines JHP conducted yeast one-hybrid screens MHL and JY prepared OX lines and...

Ngày tải lên: 11/08/2014, 11:21

... better understanding the regulatory mechanisms associated with cold-responsive genes, we may better understand the molecular differences and similarities between cold acclimation and chilling ... growth and fruit quality Authors' contributions AT: identified and cloned the promoters AT, MS, LM and HS drafted the manuscript AT and MS: performed the digital expression analysis AM and AT: ... plants, including three dehydrins (C30, C254 and C304), three chitinases (C910, C2131 and C2441), four thaumatin-like proteins (C1708, C2177, C2317 and C2147), and polygalacturo- Page 10 of 15 (page...

Ngày tải lên: 12/08/2014, 03:21

... selection between canary yellow and red watermelon [29] Thus, lycopene βcyclase plays a major role in carotenogenesis of different colored flowers and fruits Isolation and characterization of CYC-B ... promoter::reporter fusion molecular genetic screens to identify mutants impaired in CYC-B expression, and thus can be a valuable tool in understanding carotenoid metabolism in tomato Methods Isolation and cloning ... promoter has been characterized in tomato [8] Isolation and characterization of promoters of carotenoid biosynthesis pathway genes will help understand the regulation of carotenoid biosynthesis...

Ngày tải lên: 12/08/2014, 03:21

... product and authentic standard Example of comparison between absorption spectrum of the reaction product and authentic standard Absorption spectra of p-coumaroyl-quinate: standard (dotted line) and ... Science, Switzerland) by a 15–60 exposure to X-ray film Authors' contributions SL and FB planned and supervised the work CC and AH carried out the molecular genetic studies; EP and AA carried out ... mode; and the fragmentor was set at 120 eV for the standard and the isolated compounds Individual polyphenols were identified by comparison and combination of their retention time, UV-Vis and mass...

Ngày tải lên: 12/08/2014, 05:20

... White Candidate for reproduction rcp(5;8)(p12;q21) sire + one sister March 2000 French Landrace (FL) Candidate for reproduction rcp(15;17)(q24;q21) June 2000 Crossbred (3/4 FL; 1/4 Meishan) Candidate ... November 2000 Sino-European synthetic line Candidate for reproduction rcp(5;8)(p11;p23) full- and half-sibs February 2001 Large White Candidate for reproduction rcp(3;15)(q27;q13) offspring (analyses ... laboratory techniques used [27] G and/ or R banding is presently used systematically and allows the detection of rearrangements modifying very slightly the size and/ or the banding profiles of the chromosomes,...

Ngày tải lên: 14/08/2014, 13:21

... 23-30o C and 80-90% relative humidity The house dust mites primarily feed on skin scales and dander while the storage mites feed on plants and micro-organisms (Ho and Nadchatram, 1984; Andersen ... (DC) producing IL-12 and IL-18 enhance TH1 cell development, whereas DCs expressing ICOS ligand and producing IL-10 enhance the development of TH2 and TR cells TR cells and TH2 cells develop ... showing IgE binding to DF414 and DF1096; DF414 and BT796; and BT796, BT1313 and BT1694 176 IgE binding reactivity of mites FABP homologues tested by 55 Singaporean sera and 42 Italian sera that are...

Ngày tải lên: 12/09/2015, 11:08

... H and Kadowaki, K (1985a) Flocculant production by Paecilomyces sp Taxonomic studies and culture conditions for production Agric Biol Chem., Vol.49, No.11, 3151-3157 - 299 - Journal of Water and ... soil was washed times with distilled water and suspended in 1,000 ml of water The mixture was allowed to stand for 10min and the supernatant was collected and used for the flocculating test Deflocculated ... procedure - 293 - Journal of Water and Environment Technology, Vol.3, No.2, 2005 Components of extracellular polymer Components of the extracellular polymer were analyzed and the results are summarized...

Ngày tải lên: 05/09/2013, 09:08

... NRE(A+B)); and in both Box A and B, NRE(A–B–) (C) Qualitative analysis of LacZ reporter activation in the interaction of APUM-2 and APUM-7 with NRE WT and NRE mutants The iron responsive element RNA and ... human and mouse possess two; and Drosophila and Dyctiostelium have only one member [5] Recently, new studies have revealed the presence of ten, two and one homologs in Trypanosome, Plasmodium and ... positions 12, 13 and 16, similar to human and Drosophila Pumilio On the other hand, the amino acids in these positions in APUM-7 to APUM-12 are more similar to those found in yeast Puf4 and Puf5 proteins...

Ngày tải lên: 18/02/2014, 06:20