gene cloning and dna terry brown

... Basic Principles of Gene Cloning and DNA Analysis 1 Why Gene Cloning and DNA Analysis are Important Vectors for Gene Cloning: Plasmids and Bacteriophages 13 Purification of DNA from Living Cells ... Cloning and DNA Analysis in Biotechnology 223 13 14 15 16 Production of Protein from Cloned Genes 225 Gene Cloning and DNA Analysis in Medicine 245 Gene Cloning and DNA Analysis in Agriculture 264 Gene ... Principles of Gene Cloning and DNA Analysis 1 Why Gene Cloning and DNA Analysis are Important 1.1 1.2 1.3 1.4 1.5 1.6 The early development of genetics The advent of gene cloning and the polymerase...

Ngày tải lên: 06/03/2014, 22:20

... Basic Principles of Gene Cloning and DNA Analysis 1 Why Gene Cloning and DNA Analysis are Important Vectors for Gene Cloning: Plasmids and Bacteriophages 13 Purification of DNA from Living Cells ... Cloning and DNA Analysis in Biotechnology 223 13 14 15 16 Production of Protein from Cloned Genes 225 Gene Cloning and DNA Analysis in Medicine 245 Gene Cloning and DNA Analysis in Agriculture 264 Gene ... Principles of Gene Cloning and DNA Analysis 1 Why Gene Cloning and DNA Analysis are Important 1.1 1.2 1.3 1.4 1.5 1.6 The early development of genetics The advent of gene cloning and the polymerase...

Ngày tải lên: 19/03/2014, 09:20

... rspL20 mtl-1xyl-5) [20] was used for cloning of the mutated XIs using pGEMÒ-T Easy vector (Promega, Madison, WI, USA) PCR mutagenesis Random mutagenesis of the XI gene (xylA) was performed under conditions ... broader, and extended into the alkaline region, compared with the wild-type XI The wildtype showed no XI activity at pH and 10 For M-1024 and M-1026 the speci®c activity at pH and 10, was 66 and 45%, ... wild-type showed 88 and 74% of the maximum activity with Co2+ and Mg2+, respectively The mutants, on the other hand, were less activated by Co2+ and Mg2+ Kinetic properties of D-xylose and D-glucose...

Ngày tải lên: 31/03/2014, 15:20

... broad-range PCR, cloning and sequencing the entire 16S rDNA and demonstrated the presence of a large number of bacterial DNA in the synovial tissue (ST) of patients with ReA, UA, and other arthropathies ... at -80°C until analysis Automated DNA extraction from synovial fluid and broad-range PCR amplification of 16S rRNA genes, cloning and sequencing of bacterial DNA For 10 minutes, 500 μl of SF were ... RA and OA patients In addition, DNA from 20 bacterial species was detected in both the study and the control groups In ReA and UA patients, apart from Shigella sonnei DNA, there was no other DNA- derived...

Ngày tải lên: 09/08/2014, 14:22

... and Xho I restriction site was introduced on the end of the proposed synthetic trypsin inhibitor gene The sequences of the forward and reversed primers are shown on fig and of the synthetic gene ... Lebl and Richard A Houghten (Editors), American Peptide Society, 2001, pp 387-388 Bozenna Rempola, T-Wilusz, W Markiewicz and M.Fkus, Synthesis, cloning and expression in E.coli of the gene coding ... primers and the purified total MCo DNA as template The conditions for this experiment were established as: 150ng of each primer; 200àM of dNTPs ;400ng of the purified total MCo DNA Cloning of...

Ngày tải lên: 12/02/2014, 10:20

... as a genomic DNA clone were isolated and sequenced, and the expression and modulation of this molecule was studied Results Cloning and characterization of rainbow trout IL-11 A cDNA clone containing ... number AJ867256) containing the IL-11 gene and its 5¢- and 3¢-flanking region was sequenced The trout IL-11 gene has a five exon ⁄ four intron organization and all introns are phase except intron ... interleukin-11 Fig Nucleotide and deduced amino acid sequences of the trout IL-11 gene The genomic DNA (upper line, accession number 867256), cDNA (middle line, accession number AJ535687) and amino acid (lower...

Ngày tải lên: 07/03/2014, 16:20

... sensitive, and that the pA1lacO-1 promoter and the lacI q1 gene are surrounded by a DNA fragment originating from upstream the gyrA gene and a fragment containing the N-terminal part of the gyrA gene ... of 0.005 and growth was followed for at least ®ve generations before sampling All samples were withdrawn between D540 ˆ 0.2 and 0.4 Enzymes Restriction enzymes, T4 DNA ligase, and T4 DNA polymerase ... containing DNA from the gyrA locus on pGYRABTS with DNA fragments taken from upstream the topA gene and a fragment containing the N-terminal part of the topA gene pGYRABTS was ®rst digested with KpnI and...

Ngày tải lên: 18/03/2014, 01:20

... of an argon laser and an emission range between 505 and 525 nm for fluorescence detection as described previously [9] The genomic DNA sequences of the canine and murine NICN1 genes were determined ... human, dog and mouse the NICN1 gene is immediately followed by the AMT gene for aminomethyltransferase, an enzyme of glycine metabolism [4,11] The promoters of the canine and murine NICN1 genes lack ... panels (MTCTM) panels of first-strand cDNAs (BD Biosciences Clontech) The concentration of the cDNAs in the used panels is normalized to four different housekeeping genes The PCR analyses of the...

Ngày tải lên: 23/03/2014, 21:20

... 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, protein bands after renaturing ... Second-strand cDNA was amplified by 5¢ PCR primer 5¢-AAGCAGTGGTATCAACGCAGAGT-3¢ and CDS III ⁄ 3¢ PCR primer, using Advantage DNA Polymerase from Clontech Screening of cathelicidin-encoding cDNAs and ... the gene cloning and characterization of three avian cathelicidin orthologs, namely Cc-CATH precursors from Coturnix coturnix, is reported, and the relationship between quail cathelicidins and...

Ngày tải lên: 28/03/2014, 23:20

... tumor suppressor gene and is a central regulator in oncogenesis Overexpression of the Pokemon gene can decrease expression of the ARF gene, which in turn results in p53 degradation and oncogenic ... Pokemon gene both inhibited oncogene-mediated cellular transformation and induced cell senescence and apoptosis [22,26] Therefore, Pokemon plays a crucial role in cell tumorigenesis and may be ... sequences (254 and 106 bp) between the NEGU and NEG-D elements The distance between the NEG-U and NEG-D elements was 253 and 106 bp in D-254 and D-106, respectively The right-hand panel shows...

Ngày tải lên: 30/03/2014, 04:20

... kinase Screening of the venom gland cDNA library Ten thousand clones from the venom gland cDNA library were analyzed by the 32P-end-labelled oligonucleotide probe High and low density screenings of ... FEBS 2002 Molecular cloning and function of gene BmKIM (Eur J Biochem 269) 4469 Synthesis of oligonucleotide probe The oligonucleotide probe used to screen the venom gland cDNA library, constructed ... restriction enzyme site and stop codon, corresponded to positions 227–246 of BmKIM cDNA PCR was performed and the PCR product was cloned into pGEX-5X-1 after digestion with BamHI and XhoI and purification...

Ngày tải lên: 31/03/2014, 09:20

... Chitosanase Penicillium sp Gene cloning Expression The GenBank/EMBL/DDBJ accession numbers for the 18S rRNA gene and chitosanase gene of strain D-1 are JF950269 and JF950270, respectively Introduction ... still unclear, and the gene cloning and expressing are seldom reported It is of interest to characterize the gene structure of fungal chitosanase in relation to their catalytic function and compare ... Extraction of chromosomal DNA and gene cloning of chitosanase by I-PCR The chromosomal DNA of strain D-1 was extracted by the phenol-chloroform method (Chomczynski and Sacchi 1987) Based on the...

Ngày tải lên: 21/06/2014, 19:20

... procedures, and performed clinical and rheumatological data analyses AZ, CB and EC conducted assessment of Chlamydia trachomatis serology and DNA extraction BJ and JS participated in the design and coordination ... DNA in patients with ReA and UA using broad-range PCR, cloning and sequencing of almost the entire 16S rRNA gene The use of this approach revealed the identity of potential bacterial causes and ... MgCl2 and 1.25 units of Takara Ex Taq DNA polymerase (Takara Ex taq, Otsu, Shiga, Japan T4 Gene 32 Protein (5 μg/μl; USB Corp, Cleveland, Ohio) was added to the PCR mix followed by 2.5 μl of DNA...

Ngày tải lên: 09/08/2014, 10:23

... 295 to 300, and included egg weight, Haugh unit, egg index, percentage of yolk, percentage of albumen and shell strength [22] DNA isolation, RNA isolation and cDNA synthesis Genomic DNA was extracted ... population mean; Si and Gj are the fixed effects of each generation and genotype, respectively, and εij is the random error Values are considered significant at P < 0.05 and are presented as ... as least square means ± standard error Results and discussion Molecular cloning and sequence analysis of duck VLDLR Using RACE and sequence matching techniques, a cDNA sequence covering the whole...

Ngày tải lên: 14/08/2014, 13:21

... in the various gene features and expression variation are still strong for average and highly expressed genes and weak for lowly expressed genes (data not shown) Highly expressed genes also show ... (Tables and 3) Highly expressed genes also tend to have shorter gene features and fewer introns than average expressed genes, which are, in turn, shorter than lowly expressed genes (Tables and 3) ... coding regions and 5'UTRs is significantly higher in sex-specific genes than non-sex-specific genes (the pooled class of sex-biased and unbiased genes) Male-specific and male-biased genes have lower...

Ngày tải lên: 14/08/2014, 20:22

... 2.5.3 Cloning of udu cDNA constructs containing the wild type allele and the udusq1zl allele (pcDNA3.1- udu-wt, pcDNA3.1- udu-T2976A) 61 2.5.4 Cloning of Flag-tagged and HA-tagged udu cDNA ... Positional cloning and gene structure of udu gene 108 Figure 4.12 The nonsense mutation in Ensemble Gene 108 tu24 ENSDARG00000005867 (udu gene) in udu and udusq1zl mutants Figure 4.13 udu cDNA sequence ... 21,503 genes, 130 pseudogenes and 3,421 non-coding genes The zebrafish genome project greatly simplifies the process of mapping the mutant genes 1.2.4.1.2 Principles of Positional Cloning In general,...

Ngày tải lên: 14/09/2015, 17:54

Ngày tải lên: 21/09/2015, 21:25

... underlying cellular function (reviewed in Postlethwait and Talbot, 1997) The two main approaches of cloning mutated genes, positional cloning and candidate gene approach, have benefited greatly from the ... expression and function of the 29 novel human orthologous gene Materials and methods Chapter II Materials and Methods 30 Materials and methods Cloning of Full Length Zebrafish klhl cDNA 1.1 Rapid ... species would not only help us understand what makes us genetically different, it may also help us understand our genes, their regulation and expression and their complex interactions (Murphy...

Ngày tải lên: 03/10/2015, 20:57

... 3.2.4 3.2.5 Materials and Methods Cell lines and Transfection Quantitation of FAT10, p53, p21 and MDM2 and β-actin transcript Generation of siRNA constructs against p53 Generation of construct ... cancer and infectious diseases, FAT10 gene is a non-class I gene (Fan et al., 1996) Expression of FAT10 gene does not affect cell surface expression or antigen presentation of MHC class I genes ... gastrointestinal system, kidney, lung and prostate gland, but not in tissues from the brain and adrenal gland The reticuloendothelial system (e.g thymus, spleen) and the gastrointestinal system show...

Ngày tải lên: 03/10/2015, 20:57

... this journey, and I would be remiss if I not acknowledge them Thanks, mom and dad, for giving me the latitude to what I wanted to do, and generally having faith in me Your patience and generosity ... paracaspases and metacaspases Depending on the tertiary structure and sequence similarity, metacaspases can be divided into two classes Type I metacaspases are generally found in plants and fungi, and ... Molecular cloning Molecular cloning was carried out as described in section 2.1.10 30 MATERIALS & METHODS 2.3.6 DNA sequencing DNA sequencing was carried out as described previously for the PfMCA1 gene...

Ngày tải lên: 03/10/2015, 20:57