cation of a clone from a gene library

... TACAAT TATAAT TAAAAT TACTAT TATAAT TAATTT TATAAT TATAAT + – – + – + + CIRCE – This work [10] [9] [61] [62] [63] [38] [64] [37] Fig SDS/PAGE analysis of expression and puri cation of the recombinant ... alkaline phosphatase Materials and methods Bacterial identi cation and culture conditions DNA manipulation and sequence analysis Plasmid DNA preparation, puri cation of DNA from agarose gel, and restriction ... spoIIG amyR rA consensus r70 consensus TTAGACA TTAGACA TTGTACA TTGTTAC TTGTATT TTGACAG TTGTTTT TTGACA TTGACA 17 17 17 20 17 21 16 16–18 16–18 TTTG TTTG GTTG TATG TATG CTTG TGTG TNTG – TACAAT TACAAT...

Ngày tải lên: 16/03/2014, 16:20

... cluster of S alboniger [6] They were accordingly named ataP3, ataP5, ataP4, ataP10 and ataP7 The two additional ones were named ata12 and ataPKS1 (Figs and 3) All shared a codon usage and a G+C ... complementation assays The ataP5, ataP4 and ataP10 genes were also independently inserted in the pIJ702 vector and the resulting plasmids pA 2A5 , pA 2A4 and pA 2A1 0, respectively (Materials and methods), ... in A2 0 1A biosynthesis In contrast, upstream of ard1 there are six contiguous coding sequences with identical orientation (Figs and 3A) Of these, five (ataP3, ataP5, ataP4, ataP10 and ataP7) are...

Ngày tải lên: 21/02/2014, 01:21

... second set, A5 2 (5¢AGAGTGAGCAGGTAGCGAGAGGAG-3¢)/TRHR-8 (5¢-GGGGGTGTAGAGGTTTCTGGAGAC-3¢); third set, A5 3 (5¢-CGAGAGGAGCATTAGA-TAGATG Ó FEBS 2002 CAG-3¢)/TRHR-9 (5¢-GCCGAAATGTTGATGCCCA GATAC-3¢) (Fig ... CGTAACTTTTGCTG-3¢)/TRHR1-4 antisense (5¢-TC TGTTAAATGTACCTAAGTAGGCA-3¢) and TRHR2-2 sense (5¢-CAGCAAAATGGAAAATAGTAGC-3¢)/ TRHR2-4 antisense (5¢-CGACACTGTAGTAG-AGAT CACC-3¢), respectively The PCR ... 32281–32287 31 Itadani, H., Nakamura, T., Itoh, J., Iwaasa, H., Kanatani, A. , Borkowski, J., Ihara, M & Ohta, M (1998) Cloning and characterization of a new subtype of thyrotropin-releasing hormone...

Ngày tải lên: 21/02/2014, 03:20

... folded GABARAP [22] The addition of CRT to GABARAP resulted in the disappearance of GABARAP resonances, a clear indication of binding (Fig 3B) Only weak amide signals for a Gln ⁄ Asn side chain and ... et al (2001) Functional specialization of calreticulin domains J Cell Biol 154, 961–972 Harada K, Takeuchi H, Oike M, Matsuda M, Kanematsu T, Yagisawa H, Nakayama K-II, Maeda K, Erneux C & Hirata ... interact with CRT By contrast with GABARAP, no quantitative data are available for these interactions Most importantly, the interaction of CRT with GABARAP occurs with natively folded GABARAP This...

Ngày tải lên: 16/03/2014, 05:20

... Muroya A, Tsuchiya D, Ishikawa M, Haruki M, Morikawa M, Kanaya S & Morikawa K (2001) Catalytic center of an archaeal type Ribonuclease H as revealed by X-ray crystallographic and mutational analyses ... Structure of ribonuclease H phased at A resolution by MAD analysis of the selenomethionyl protein Science 249, 1398–1405 Ishikawa K, Okumura M, Katayanagi K, Kimura S, Kanaya S, Nakamura H & Morikawa ... 5¢-TTCAAGAATTCTCATGTTTTGAC -3¢ for the 5¢-primer, 5¢-CGCGTCGACACTAACAG GGCTGATTGACGAGTC-3¢ for the 3¢-primer, 5¢TCTACCAGAG ATG TCGACATTATCGGTTGTG-3¢ for the 5¢-fusion primer and 5¢-TAATGTCGA CAT CTCT GGTAGACTTCCTGTAA-3¢...

Ngày tải lên: 16/03/2014, 13:20

... according to the manufacturer’s instructions The cDNA was amplified with Lipases in papaya latex the primers 5¢-ACTCAAATCACTAGTATTCTTCCACCA-3¢ and 5¢-CATTTGAACATAAACATGAACAAATAAGTT-3¢ and the following ... percentage of release from position was calculated as follows: area fatty acid ⁄ (area fatty acid + area lysoPtdCho) Pancreatic PLA2 was used as a positive control, and T lanuginosus lipase as a PLA1 ... close relative of C papaya is V heilbornii (mountain papaya or babaco), formerly known as Carica pentagona [30] The Vasconcellea and Carica genera are close enough that hybrids can be obtained under...

Ngày tải lên: 22/03/2014, 17:20

... 5¢-TATGTAGGAGCTACAAGAAACATTGAAGGC-3¢ 5¢-CAATTCTTACTAGCTGCAGGGAGGGTTGGT-3¢ 5¢-GAATTGGCCCTAATTTGCACATTAAAGATA-3¢ 5¢-GCCTGAAGTTTTTGGTTCACAGTGAAATTT-3¢ 5¢-ACACTCTGAAGGGGAATTAGGATTAAGAAA-3¢ 5¢-CTGACTATACAAGGCTGTGTGCCACAAAAC-3¢ ... 5¢-CTGACTATACAAGGCTGTGTGCCACAAAAC-3¢ 5¢-GAAAACACATCACGTACCCTCCCTCTCTGCG-3¢ 5¢-ACATCATGGTATATTATTCATTTAGCTGACACTTT-3¢ 5¢-ATGGATGAGGAAATCGCTGCCCTGGTC-3¢ 5¢-TTAGAAGCACTTCCTGTGAACGATGGA-3¢ 3835 A novel ... ATGAGCCGGAGAACAAGCGAAACT-3¢) and antisense (5¢-CGCGGATCCTTATATAGTGAAGCGTATTGGAAG FEBS Journal 272 (2005) 3828–3837 ª 2005 FEBS S Yasuda et al A novel zebrafish cytosolic sulfotransferase AGGACA-3¢)...

Ngày tải lên: 23/03/2014, 15:20

... recombinant NaoA after Sephadex G75 chromatography; lane 5, purified recombinant NaoA after DEAE-Sepharose Fast Flow chromatography; lane 6, standard molecular mass markers (phosphorylase b, 97 kDa; ... this gene was designated naoA (nitroalkane-oxidizing enzyme gene) Expression of naoA in E coli To study the function of the naoA gene, it is necessary to obtain an adequate amount of NaoA protein ... metal ions may also act in other ways We conclude that NaoA is a nitroalkane dioxygenase-like enzyme rather than a nitroalkane oxidase Its characteristics are not identical with those of any...

Ngày tải lên: 31/03/2014, 08:20

... primers A3 were as follows: 5¢-GCCGGATCCCCGATGACGATGACAAG GATGGATATATAAGA-3¢ as forward primer containing a BamHI restriction enzyme site (underlined) and corresponding to five codons encoding an enterokinase ... such as A (Ala) or D (Asp) Only AaHIT4 and BmKAS, a specific anti-insect toxin, also contained a Tyr residue at this position AaHIT4, the unique anti-insect toxin also has a toxic effect on mammals ... These data indicated that rBmKIM had toxicity to both mammals and insects, though the toxicity was at a lower level Assay of antiarrhythmia activity Table illustrates the effects of rBmKIM on aconitineinduced...

Ngày tải lên: 31/03/2014, 09:20

... TCCACCACCCTGTTGCTGTA CCCTTTGCCATTGTTATA TCAGAGGTCGGTCAAACG ACGGGCAATACCCTCTGG AGGACTCGTCGGGAAGCA CCACCAGTCCTGATGCCA AGCCTTCAAGCGGGGAG CAAGCTCTTGAAAGGCATCG AGAGGGCTTCCTGGCTCA AACGCAGAGTACGCGGG CAGCCAAGGGTGGTCCTGC ... ACCACAAGTCCATGCCATCAC TCCACCACCCTGTTGCTGTA TCTGGGAGGTGTCAACGAG GGTCTTGGCATAGTCATAGGT GCTACGCGTTCAGCACTATCAGATCGGG GCTACGCGTCCTTCCTAGCCCCACCT GCCACGCGTATCTATTAGCCTCGTCCCAC GATACGCGTCAGCCCGCCACATCTCAG GATACGCGCATAGCCCAGCCCATCTC ... CAGAGTACTTTTTTTTTTTTTTTT AGCAGGGGCTGTATCGTC NGTCGASWGANAWGAA GTNCGASWCANAWGTT WGTGNAGWANCANAGA NCAGCTWSCTNTSCTT CTGCTCTCTACGAAAGGCCGTGC CCCAACTCGCCGCCTACCTTCC TCGCCGCCTACCTTCCGGAGCGC ACCACAAGTCCATGCCATCAC...

Ngày tải lên: 14/08/2014, 13:21

... and µL was used to inoculate a new overnight culture This was repeated on day 2, 3, 4, and until 100 generations had passed BAC DNA from each clone was prepared as above on day and NotI and HindIII ... libraries, Genomics 52 (1998) 1–8 [18] Rogel-Gaillard C., Bourgeaux N., Billault A. , Vaiman M., Chardon P., Construction of a swine BAC library: application to the characterization and mapping of ... Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector, Proc Natl Acad Sci USA 89 (1992) 8794–8797 [21] Suzuki K., Asakawa S.,...

Ngày tải lên: 14/08/2014, 13:22

... gtgactgcag gaaggggatc cggctgctaa caaagcccga 280 281 aaggaagctg agttggctgc tgccaccgct gagcaataac 320 321 tagcataccc 361 TTTTTGCTGA cttggggcct AAGGAGGAAC ctaaacgggt TATATCCGGA cttgaggggt TATCCCGCAA ... MCoTI-II gene cttaaggtatactcgccgtcgcgtaccgccgcacacgggcttt gaattccatatgagcggcagcgatggcggcgtgtgcccgaaa M S G S D G G V C P K taagacttttttacggcggcgctatcgctaacgggcccgcgc attctgaaaaaatgccgccgcgatagcgattgcccgggcgcg ... Gtttttgctt ggatcccagg ttgttgtaca gaatgctggt 120 121 catatgagcg gcagcgatgg cggcgtgtgc cgaaaattct 160 161 gaaaaaatgc cgccgcgata gcgattgccc gggcgcgtca 200 201 tttgccgcgg caacggctat tgcggctaac tcgagcccgg 240...

Ngày tải lên: 12/02/2014, 10:20

... TCATCAT-3¢; rOMM-64-IV, 5¢-CGCGGATCCGCCCCT GTTAATGATGGAACC-3¢ and 5¢-CGCCTCGAGCTAA GAAGACTGGGCTGCCAG-3¢; rOMM-64-V, 5¢-CGCGG ATCCAGGCAAGATTTTAAGCATCCA-3¢ and 5¢-CGCC TCCACCTAAGAGGCATCCTTGTCCAC-3¢; ... 5¢-CGCCTCCA CCTAAGAGGCATCCTTGTCCAC-3¢; rOMM-64-II, 5¢CGCGGATCCACCGTAGACACTTATGATATA-3¢ and 5¢-CGCCTCGAGCTAAGAGTCAGCTTGCACGTC-3¢; rOMM-64-III, 5¢-CGCGGATCCGCTGATGTGACCAGT GATGAC-3¢ and 5¢-CGCCTCGAGCTATTTGGGCTCTT ... digestion of genomic DNA contamination in the total RNA was confirmed by lack of ampli cation of a b-actin mRNA fragment by PCR using a pair of primers (5¢-ATCACCATCGGCAACGAGAG-3¢ and 5¢-TGGAGTTGTAGGTGGTCTCGTG-3¢)...

Ngày tải lên: 18/02/2014, 17:20

... N-terminus of P40 as it appears in protein extracts of M pneumoniae Determination of the N-terminus of P40 by Edman degradation failed due to the limited amount of starting material An alternative ... processing of the gene product of MPN 142 Cleavage into P40 and P90 takes place after amino acid 454 The N-terminus of the mature P40 starts with amino acid 26 The molecular mass of about 36 000 Da of ... FEBS I Catrein et al nated sulfonic acid group and lost during fragmentation, which causes neutralization of the b-fragments The formation of sodium and potassium adducts was possibly a general property...

Ngày tải lên: 19/02/2014, 18:20

... glycoforms were also observed for strains 425/93 and 1000 (Table 1) Core oligosaccharide from the 1000 galE mutant strain was also prepared and examined by MS A range of molecular masses was found ... presence of an additional phosphate group (1032), and additional PEtn moiety (1075), an additional phosphate group and an additional PEtn moiety (1155) and one additional phosphate group and two additional ... liberated oligosaccharide and an associated extracellular polysaccharide Carbohydr Res 279, 227– 244 24 Aspinall, G.O., Monteiro, M .A & Pang, H (1995) Lipo-oligosaccharide of Campylobacter lari...

Ngày tải lên: 08/03/2014, 02:20

... percent of the amount of NADH used in our standard NR assay, and 60% of the amount of ATP in a GS assay, was converted into AMP within 10 at 30 °C by an amount of a Mono Q fraction that appeared ... in a 100-lL incubation at 30 °C for 10 min, using the amounts and concentrations of ATP and NADH used in standard GS and NR assays, respectively Data are presented as mean ± SEM Cofactor AMP generated ... trichloroacetic acid The A5 04 was measured and the c-glutamyl hydroxamate produced was quantified using commercial c-glutamyl hydroxamate as standard Control assays were performed in the absence of ATP...

Ngày tải lên: 08/03/2014, 08:20

... the appearance of a significant deacylation activity Any bacterial contamination of our protein preparations that might be responsible for activation of the PLB precursor at prolonged storage was ... staining as described in Materials and methods Marker proteins and their corresponding molecular masss are indicated in lane Lane 2, material from the acid extracts of granules; lane 3, material ... and bacterial, protease contamination Protein concentration was determined with a Bio-Rad protein assay kit using BSA as a standard according to the manufacturer’s protocol Any bacterial and protease...

Ngày tải lên: 16/03/2014, 04:20

... GGGCAAGCTTGCgGCcGCAATCTGCTTTCGAcAGAATCTGAAcACATAttcgAAAAAGTATATGC GGGCAAGCTTGCgGCcGCAATCTGCTTCCGAcAGAATCTGAAcACATACAgCTATATATATAGG GGAATTCTGCAGTTAGCATTTgagCTCtccCTTGCACAAgAAGTCCGG Ó FEBS 2003 3020 P Prijatelj ... GGGCaagcTTCCCCGTCTCCtCCAGGATCATCtTCCCG GGGCAAgcttgCTaTTcCCTCCTACTCCTcTTACGGATGCTACTGCGGCtgGGGGGG GACTGCAaCCCCAAAtCGGACAG CAAATACaAgCGGGtGAACGGGGCTATCGTCTGTGaAAAAGGC GGAATTCGCgGCcGCCtTGTCACACTCACAAATCCGATTCTC GGGCAAGCTTGCgGCcGCAATCTGCTTTCGAcAGAATCTGAAcACATAttcgAAAAAGTATATGC ... each ampli cation primer, 400 lM of each of the four deoxyribonucleoside triphosphates, Taq DNA ligase buffer (New England BioLabs) and 100 ng of template DNA Manual hot start ampli cation was...

Ngày tải lên: 17/03/2014, 03:20

... GAATTCAGAGAAAGAGATACGAGATGGA-3¢) and U660033Not (5¢-TCATAAGGCGGCCGCCTACATCG ATCTTAATCTGCTCAA-3¢) A fragment carrying At3g21260 cDNA was amplifed from A thaliana RNA by RT-PCR RNA was isolated from ... GGTTTCTAAACCAACACGT-3¢) and GLTP1PRONBAM (5¢-CTCCTTGGATCCGCCTGAGAATTGAAAAA GGTGGG-3¢) A 1.3 kb fragment carrying the At1g21360 promoter was amplified using primers 21360PRUXBA (5¢-AACGATCTAGATTAAGAATGTAATCACATTAGG ... Histochemical GUS activity assays A 1.8 kb DNA fragment carrying the At2g33470 promoter was amplified from the A thaliana Col-0 genome using primers GLTP1PROUXBA (5¢-AGACTGCTCTAGAATG GGTTTCTAAACCAACACGT-3¢)...

Ngày tải lên: 23/03/2014, 07:20

... CTTTATATTCATCCAGTGGCTGTATTCTGTGGGCCACACCAG CTGGTGTGGGCAGCAGAACTCAGCCATTGGATGAATATAAAG CTTTATATTCATCCAATGGCTGAGTTCTGCTGCCCACACCAG CTGGTGTGGGCAGCAGAATACAGCCATTGGATGAATATAAAG CTTTATATTCATCCAATGGCTGTATTCTGCTGCCCACACCAG ... Sense Antisense Sense Antisense Sense Antisense Sense Antisense CTGGTGTGGCCCACAGAACTCAGCCACTGGATGAATATAAAG CTTTATATTCATCCAGTGGCTGAGTTCTGTGGGCCACACCAG CTGGTGTGGCCCACAGAATACAGCCACTGGATGAATATAAAG CTTTATATTCATCCAGTGGCTGTATTCTGTGGGCCACACCAG ... LumiGLOTM was from Cell Signaling (Beverly, MA, USA), and AdvantageÒ polymerase mix was from Clontech (Palo Alto, CA, USA) All other reagents were of the best quality and commercially available Expression...

Ngày tải lên: 23/03/2014, 09:21