chitosan conjugated dna nanoparticle delivery systems for gene therapy

... 45 s, annealing at 60 °C for 30 s, extension at 72 °C for min, Cloning of CrPrx cDNA and gene and a final extension at 72 °C for 10 The corresponding genomic sequence for CrPrx was PCR-amplified ... manutions of the cDNA sequence, respectively, for subcloning facturer’s instruction, and used as the template for PCRs the CrPrx gene PCR amplifications were performed with degenerate oligonucleotide ... et al Table References used for sequence and expression data pre42 sented in Fig for phylogenetic analysis NA, not available A Label B Fig Intron mapping of CrPrx gene (A) Lanes M show size markers...

Ngày tải lên: 23/03/2014, 09:21

... Standard protocols were used for the transfer of RNA and DNA after electrophoretic separation [24] Hybridization of RNA or DNA transferred to nylon membranes was performed using a nonradioactive ... full-length cDNA Two degenerated primers were designed according to previously determined peptides Sequences were YTTRTC RCANACNACRTANACNCKRTGNACNSWNCCRTA for MJErev4 and GAYATGGCNGCNWSNGGNATH AAYCC for ... homologous primers were designed for the rapid amplification of cDNA ends (RACE) (RACEfor: GTGA CAGCTTTCATGCCTGG; and RACErev: ATCCTGT CCGTTGTTGTAAAC) 5¢- and 3¢-RACE was performed using the SMART II...

Ngày tải lên: 16/03/2014, 18:20

... transcribed into cDNA in 20 lL reaction mixture using SuperScript II RNase H-Reverse Transcriptase (Invitrogen) The generated cDNA was used as template for PCR, which was performed with 1.5 mm ... 3¢-rapid amplification of cDNA ends technique (RACE) To obtain the 3¢-terminal of CaLP cDNA ends, the initial round of PCR reaction was conducted with a gene- specific forward primer LS11 (5¢-TCTCGTGGAAGAAATCGACA-3¢) ... before hyde overnight Digoxigenin-labeled RNA probes were generated from the cDNA clone encoding oyster CaLP in plasmid using a DIG RNA Labeling kit (Roche), with T7 and SP6 RNA polymerase for...

Ngày tải lên: 16/03/2014, 23:20

... (PE Biosystems) on an ABI 310 Genetic Analyser (PerkinElmer) Complete nucleotide sequences were determined for both strands of the cDNAs and analysed by the DNASTAR program package (Lasergene) ... oligo(dT) cellulose (Boehringer) A directional cDNA library was constructed using lg poly(A)+ RNA according to the instructions for a Stratagene cDNA synthesis kit (Uni-ZAP XR) The resultant library ... detected for cuts by BamHI or HindIII enzymes while up to 10 bands were found when genomic DNA was digested with EcoRI (Fig 3) These results indicate that a small multigene family of at most five genes...

Ngày tải lên: 18/03/2014, 01:20

... kb PCR fragment that was used for the Northern blot probes RESULTS Isolation and sequence of dog and human p76RBE cDNA Dog p76RBE cDNA was isolated in a search for genes whose expression is regulated ... Netherlands), pcDNA3.HA and pEGFP-C3 (Clontech, Erembodegem, Belgium) Likewise, we fused full-length keratin cDNA to the His-tag of pcDNA3 (Invitrogen, Merelbeke, Belgium) The pcDNA3.myc– RhoB ... here Dr M Spaargaren and Dr J Camonis for their kind gift of cDNAs We want to thank Patricia Cornet for the kind gift of uterine mRNAs and Vanessa Van Vooren for human thyroid RNA preparation We...

Ngày tải lên: 23/03/2014, 21:20

... products of genes, in the form of cDNAs, as a prelude to genomic DNA sequencing The rationale for this was that it would be more useful and cost effective as the protein-coding regions of our genes ... to gene Moreover, as mutagenesis screens relies heavily on “phenotype first” approach, genes with subtle lossof-function phenotypes or genes whose function can be compensated for by other genes ... identified genes and proteins in humans and other organisms This approach, while effective in identifying genes, cannot differentiate between a functional or nonfunctional (pseudogene) gene A pseudogene...

Ngày tải lên: 03/10/2015, 20:57

... performed in triplicates Amplification conditions were 40 cycles with the following profile: 95 °C for 30 s, 60 °C for 30 s, and 72 °C for For each kind of tissue, standard curves were generated for ... (Applied Biosystems, Foster City, CA, USA) 18S rRNA transcript was chosen as a reference gene for the normalization of expression data and was amplified with the 18 h and 18L primers [37] For amplifications, ... amplification reaches a fluorescence threshold after 8.49 ± 2.68 cycles for branchial plume cDNA and after 17.80 ± 4.02 cycles for trophosome cDNA (Fig 1) Similarly, RpCAtr amplification reaches a fluorescence...

Ngày tải lên: 18/02/2014, 16:20

... at 4000 g for 15 at 20 °C Additional supernatant was removed from the cell debris by ultra centrifugation at 60 000 g for 60 at °C The resulting extracellular fraction (EC) was used for the assay ... 2003 2356 Y Otsuka et al (Eur J Biochem 270) Fig Phylogenetic tree of 2BW-1 based on 18S rDNA sequence comparisons of sequences of 18S rDNA and drawn using software The numbers on some branches ... ethers: Lignin model substrates for the possible fluorometric assay of b-etherases Holzforschung 33, 134–135 16 Peizer, L.R & Vogel, H (1969) An autoclavable medium for the isolation of mycobacteria...

Ngày tải lên: 17/03/2014, 03:20

... v) formic acid (solvent C) and 0.1% (v ⁄ v) formic acid in 90% (v ⁄ v) CH3CN (solvent D), using a program of 5% (v ⁄ v) solvent D for min, a gradient of 1.05% per for 90 and 100% solvent D for ... (http://www.fasta.genome.ad.jp/) DNA constructs ISP36 cDNA clones for protein expression were prepared using the full-length human ISP36 cDNA sequence A human ISP36 cDNA clone in pSPORT1 (clone ID: ... (1 U) The suspension was treated with DNase I and RNase A (final concentration of 125 lgÆmL)1 for each enzyme) at °C for h and then centrifuged at 800 g for 15 The supernatant was designated the...

Ngày tải lên: 30/03/2014, 20:20

... task, B1 dwelled at the character's location for 250 ms to select it; this was the method B1 regularly used for typing for more than seven years For the second task, B1 raised his eyebrow to ... MMG data before generating a switch response For the disabled participant, B1, although the time taken to complete the typing task with the eyebrow was only slightly less than that for the 250 ... 10 for cued response tests, was not fatiguing to use for prolonged periods, and required minimal effort to control These results suggest that MMG may be used as a noninvasive access pathway for...

Ngày tải lên: 19/06/2014, 08:20

... the user's physical comfort as well as social comfort Traditional sensing technologies are rarely designed for continuous, on-body use: those that require skin contact are generally designed to ... are generally designed for use in stationary devices Consequently, the achievement of certain design goals for existing sensors (such as durability) is ultimately detrimental to the user's comfort ... versus 2–12 cm2 for the other sensors and so the response time for the shoulderblade foam sensor would be slightly slower than that for the other sensor positions, e.g seconds for shoulder lift...

Ngày tải lên: 19/06/2014, 10:20

... [http://www.ch.embnet.org/software/TCof fee.html] 24 HGNC Gene Family Nomenclature [http://www .gene. ucl.ac.uk/ nomenclature/genefamily/interleukins.html] 25 National Center for Biotechnology Information(Bethesda, MD, USA) ... search engine [12,13] The National Center for Biotechnology Information (NCBI) (Bethesda, MD, USA) non-redundant human database was used in both cases cDNA cloning and expression constructs A full-length ... in a GST-c19orf10 fusion gene Restriction enzyme analysis and DNA sequencing were used to confirm the fidelities of the plasmids The expression constructs were then transformed into Escherichia...

Ngày tải lên: 09/08/2014, 10:20

... to study HQT gene in C cardunculus Primer Sequence (5'-3') CODhqtFor CODhqtRev HQT -For HQT-Rev HCT-ForRT HCT-RevRT HQT-ForRT HQT-RevRT ACT-ForRT ACT-RevRT HCT -For HCT-Rev HCT-InnerFor HCT-InnerRev ... of initiating comparative QTL mapping Within gene markers, such as the ones described here for the HCT and HQT genes, are particularly suitable for general mapping, and should prove useful as ... target genes The cDNAs were performed in triplicate for each sample in 20 μl Reaction mixes contained 2× iQ SYBR Green Supermix (Bio-Rad Laboratories, USA), specific primers at 300 nM, and μl of cDNA...

Ngày tải lên: 12/08/2014, 03:20

... co-transformed into the yeast competent cells (Table 3) Three independent clones from each co-transformation were analysed for the activation of the β-galactosidase (β-gal) reporter genes As ... mentioned in Table The NA gene carried the recognition sites for EcoRI and XhoI whereas the HAt gene carried the recognition sites KpnI and XhoI restriction enzymes in their forward and reverse primers ... the P3 protein The P1 gene carried the recognition sites for EcoRI and XhoI restriction enzymes in its forward and reverse primers respectively The amplified HAt and NA genes were ligated into...

Ngày tải lên: 12/08/2014, 04:21

... Ks ofgene/Kset.pairs in the NEW gene set Distribution of distribution gene set Mouse FAM 2gene set NEW gene set Provided gene set FAM2 the gene HumanALL the mouse NEWofgeneset Additional set geneset.ALL ... provides the mouse NEW gene set Additional data file provides the distribution of Ka/Ks to Ks of the gene pairs in the NEW gene set Click geneforgeneset.s toFAM2geneset Ks of the gene pairs in the ... absolute rates For the two-copy gene families, we used the slopes for the NEW gene sets directly, whereas for the entire genome we used the two slopes of the linear functions for the ALL gene sets...

Ngày tải lên: 14/08/2014, 08:20

... a potential therapeutic for the treatment of SCI For even longer, spinal cord and brain injuries were considered inoperable and unable to regenerate following damage Regeneration studies, beginning ... this therapy in treating SCI Fortunately, great effort has been exerted to surmount these obstacles Compared to peripheral nerve grafts, advantages of using purified SCs includes the potential for ... subcutaneously for hydration and were placed in temperature controlled housing overnight for monitoring recovery All surgical and animal handling procedures were performed as approved under the Guide for...

Ngày tải lên: 24/08/2014, 09:14

... treated with the O/PVP/SDS nanoparticles at the concentration of IC50 for 72 h 89 Chapter 5, Figure The scanning results of hybridizing signals on gene chips displaying the gene expression alteration: ... of chemotherapy, developed an organic arsenical, Salvarsan (Arsphenamine), which was effective in treating tuberculosis and syphilis Arsphenamine was the standard therapy for syphilis for nearly ... generates a fusion between the PML gene and the RARα gene, which encodes a transcription factor The resulting PML/RARα fusion protein blocks the expression of genes required for normal myeloid differentiation...

Ngày tải lên: 11/09/2015, 16:03

... process makes it happen for most cases After the morphogenesis reaches its final state, there is no further morphogenic diving force for cellular activity leading to morphogenesis The final state ... adhesive The interactive forces generated between cells are stronger than the tethering forces from ASGPR-galactose binding, therefore, the cells tend to support each other and form spheroids, since ... liver systems to sustain liver patient's lives before liver transplantation, establishing in vitro hepatocyte-based model 3) establishing culture model for drug metabolism/toxicity screening for...

Ngày tải lên: 12/09/2015, 08:19

... Ms Gong lingli for doing the echocardiography for the patients Thanks to Ms Connie Tse, for helping to collect the blood samples and the clinical data Thanks to Dr Sivalingam SP for his humor, ... Singapore General Hospital for their help and friendship ii Thanks to all staff of Department of Clinical Research, Singapore General Hospital, Dr Aw, Ms Cindy Goh, and the other staff Thanks for their ... Singapore for offering me the Research Scholarship and Singapore General Hospital for providing me technical and research support Finally, I would like to give the greatest gratitude to my family for...

Ngày tải lên: 14/09/2015, 12:42