scholarly journals Construction of an Expression Vector Containing Mouse-Rat Chimeric Genes Encoding a Therapeutic Antibody against CD81

2019 ◽  
Vol 28 (3) ◽  
pp. 239-244
Author(s):  
Tsutomu Yamasaki ◽  
Yuji Arai ◽  
Shuji Nakagawa ◽  
Toshikazu Kubo ◽  
Tohru Nakanishi
Keyword(s):  
Expression Vector ◽  
Therapeutic Antibody ◽  
Chimeric Genes ◽  
Genes Encoding

scholarly journals Identification of the TMV Replicase Sequence That Activates the N Gene-Mediated Hypersensitive Response

1997 ◽  
Vol 10 (6) ◽  
pp. 709-715 ◽  
Author(s):  
Hal S. Padgett ◽  
Yuichiro Watanabe ◽  
Roger N. Beachy
Keyword(s):  
Hypersensitive Response ◽  
Expression Vector ◽  
N Gene ◽  
Replicase Gene ◽  
Response Mechanism ◽  
Replicase Protein ◽  
Gene Response ◽  
Genes Encoding ◽  
Virus Genomes ◽  
High Degree

The N gene-mediated hypersensitive response (HR) in tobacco provides a high degree of resistance against most tobamoviruses by halting the progress of infection at the site of inoculation. A previous report indicated a role for the 126/183-kDa replicase in induction of the HR in tobacco containing the N gene (H. S. Padgett and R. N. Beachy, Plant Cell 5:577-586, 1993). Chimeric virus genomes were constructed in which the genes encoding the 126/183-kDa proteins of the HR-eliciting pathogen, tobacco mosaic virus (TMV), and the resistance breaking tobamovirus, Ob, were exchanged. Inoculation of the chimeric viruses to leaves of Nicotiana tabacum cv. Xanthi NN confirmed that either the replicase protein of TMV or its mRNA was responsible for induction of HR. An expression vector based on the Ob virus was used to express fragments of various replicase genes. With this approach, it was determined that the HR is caused by a portion of the replicase protein extending from amino acid 692 to 1116. Consistent with this result, Ob mutants that induce the HR on NN tobacco were found to carry mutations within the same portion of the replicase gene. The N gene-mediated HR is inactive at high temperatures, yet these mutants were able to overcome the HR at significantly lower temperatures than could TMV, indicating that the temperature sensitivity of the N gene response is manifested at the level of interaction between the virus and the defense response mechanism.

scholarly journals Nisin-Controlled Production of Pediocin PA-1 and Colicin V in Nisin- and Non-Nisin-Producing Lactococcus lactis Strains

2004 ◽  
Vol 70 (8) ◽  
pp. 5030-5032 ◽  
Author(s):  
Nikki Horn ◽  
Antonio Fernández ◽  
Helen M. Dodd ◽  
Michael J. Gasson ◽  
Juan M. Rodríguez
Keyword(s):  
Lactococcus Lactis ◽  
Chimeric Genes ◽  
Two Component ◽  
Genes Encoding ◽  
Colicin V ◽  
Nisin Producer

ABSTRACT The introduction of chimeric genes encoding the fusion leader of lactococcin A-propediocin PA-1 or procolicin V under the control of the inducible nisA promoter and the lactococcin A-dedicated secretion genes (lcnCD) into Lactococcus lactis strains, including a nisin producer, expressing the two component regulator NisRK led to the production or pediocin PA-1 or colicin V, respectively.

scholarly journals Intrachromosomal Recombination within thevsp Locus of Mycoplasma bovis Generates a Chimeric Variable Surface Lipoprotein Antigen

2001 ◽  
Vol 69 (6) ◽  
pp. 3703-3712 ◽  
Author(s):  
Inessa Lysnyansky ◽  
Yael Ron ◽  
Konrad Sachse ◽  
David Yogev
Keyword(s):  
Recombination Event ◽  
Phenotypic Switching ◽  
Upstream Region ◽  
Mycoplasma Bovis ◽  
Gene Repertoire ◽  
Genomic Locus ◽  
Chimeric Genes ◽  
C Terminus ◽  
Variable Surface ◽  
Genes Encoding

ABSTRACT A family of 13 related but divergent vsp genes was recently found in the chromosome of the bovine pathogenMycoplasma bovis. The vsp genomic locus was shown to undergo high-frequency rearrangements and to mediate phenotypic switching of variable lipoprotein antigens (Vsps) on the mycoplasma cell surface. Here we report that the vsp gene repertoire is subject to changes. Genetic analysis of M. bovis clonal isolates displaying distinct Vsp phenotypes showed that an intergenic recombination event between two closely related members of the vsp gene family, the formerly expressedvspA gene and the vspO gene, led to the formation of a new chimeric and functional vsp gene,vspC. The 5′ end of the recombination event was identified within the highly conserved vsp-upstream region, while the 3′ end was localized within the first repetitive domain (RA1) present in both vspA and vspOstructural genes. As a result, the vspC gene is an embodiment of the following domains: an N-terminus-encoding region linked to the highly conserved vsp-upstream region provided by the vspO gene; and a C-terminus-encoding region and the more distal and divergent vsp-upstream region acquired from the vspA gene. The generation of chimeric genes encoding surface antigens may provide an important element of genetic variation and an additional source of antigenic diversification within the mycoplasma population.

Expression of the Genes Encoding Ectoine Synthases from Cobetia marina CICC10367 in E. coli BL21

2012 ◽  
Vol 573-574 ◽  
pp. 1112-1116
Author(s):  
Qing Chen ◽  
Ling Hua Zhang ◽  
Ying Ying Liu ◽  
Ya Jun Lang
Keyword(s):  
Escherichia Coli ◽  
Expression Vector ◽  
Monosodium Glutamate ◽  
Induction Method ◽  
E Coli ◽  
Recombinant Vector ◽  
Medium Components ◽  
Cobetia Marina ◽  
Recombinant Type ◽  
Genes Encoding

In order to investigate the effects of the concentration of NaCl, the medium components and the recombinant type on the expression of the ectoine synthase genes ectABC in Escherichia coli BL21, ectABC and their promotor sequence from Cobetia marina CICC10367 were cloned. The cloned sequence was restructured with expression vector pET43.1 and then the recombinant vector was transformed into E. coli BL21. Two types of recombinant were obtained including recombinant ect03, the restructured ectABC and recombinant pect21, the restructured ectABC with promotor. The expression products of these two recombinants were identified by 1H-NMR and the effect of induction method and condition on the expression of ectoine was investigated. The result indicated that low concentration of NaCl was conducive to the expression of ectoine in recombinant ect03. The highest concentration of ectoine from recombinant ect03 was 216.1 g/L in MMG medium under 0.5% NaCl. The use of monosodium glutamate improved the expression of ectoine significantly. Higher concentration of NaCl (3%) was conducive to the expression of ectoine in recombinant pect21.

Molecular characterization of fatty acid α-hydroperoxide-forming enzyme (α-oxygenase) in rice plants

2000 ◽  
Vol 28 (6) ◽  
pp. 765-768 ◽  
Author(s):  
T. Koeduka ◽  
K. Matsui ◽  
Y. Akakabe ◽  
T. Kajiwara
Keyword(s):  
Escherichia Coli ◽  
Arabidopsis Thaliana ◽  
Fatty Acid ◽  
Expression Vector ◽  
Oxygen Electrode ◽  
Vector System ◽  
High Similarity ◽  
Rice Plants ◽  
Genes Encoding

Genes encoding an α-oxygenase, in Nicotiana tabacum and Arabidopsis thaliana have been recently isolated. However, the reaction mechanism of the enzyme has not so far been elucidated. In this study, a cDNA encoding the fatty acid α-oxygenase gene in rice plants was isolated. The deduced amino acid sequence showed high similarity (63.6%) to that of N. tabacum. The gene was cloned into an expression vector system, pQE-30, and expressed in Escherichia coli as a host cell. Palmitic acid as a substrate was incubated with the lysate of the cells, and the products were analysed by HPLC. A compound formed predominantly by the recombinant enzyme was shown to be n-pentadecanal. By incubating the mixture at 0 °C, 2-hydroperoxypalmitic acid was detected as a primary product and little formation of n-pentadecanal was detected. Furthermore, uptake of molecular oxygen was observed with an oxygen electrode. This indicated that the gene in rice plants encodes the α-oxygenase.

Expression of bacteriophage ϕEa1h lysozyme in Escherichia coli and its activity in growth inhibition of Erwinia amylovora

Microbiology ◽  
2004 ◽  
Vol 150 (8) ◽  
pp. 2707-2714 ◽  
Author(s):  
Won-Sik Kim ◽  
Heike Salm ◽  
Klaus Geider
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Expression Vector ◽  
Erwinia Amylovora ◽  
Kanamycin Resistance ◽  
Target Cells ◽  
Sequence Alignments ◽  
E Coli ◽  
Kanamycin Resistance Cassette ◽  
Genes Encoding

A 3·3 kb fragment from Erwinia amylovora phage ϕEa1h in plasmid pJH94 was previously characterized and found to contain an exopolysaccharide depolymerase (dpo) gene and two additional ORFs encoding 178 and 119 amino acids. ORF178 (lyz) and ORF119 (hol) were found to overlap by 19 bp and they resembled genes encoding lysozymes and holins. In nucleotide sequence alignments, lyz had structurally conserved regions with residues important for lysozyme function. The lyz gene was cloned into an expression vector and expressed in Escherichia coli. Active lysozyme was detected only when E. coli cells with the lyz gene and a kanamycin-resistance cassette were grown in the presence of kanamycin. Growth of Erw. amylovora was inhibited after addition of enzyme exceeding a threshold for lysozyme to target cells. When immature pears were soaked in lysates of induced cells, symptoms such as ooze formation and necrosis were retarded or inhibited after inoculation with Erw. amylovora.

Three-Dimensional Structure of Cloned T3 Connector Protein at 1.6nm Resolution

1990 ◽  
Vol 48 (1) ◽  
pp. 282-283
Author(s):  
José L. Carrascosa ◽  
José M. Valpuesta ◽  
Hisao Fujisawa
Keyword(s):  
Dna Sequences ◽  
Expression Vector ◽  
Three Dimensional ◽  
Deae Cellulose ◽  
Dna Packaging ◽  
Dimensional Structure ◽  
Two Dimensional ◽  
Freezing And Thawing ◽  
Three Dimensional Structure ◽  
Dimensional Reconstruction

The head to tail connector of bacteriophages plays a fundamental role in the assembly of viral heads and DNA packaging. In spite of the absence of sequence homology, the structure of connectors from different viruses (T4, Ø29, T3, P22, etc) share common morphological features, that are most clearly revealed in their three-dimensional structure. We have studied the three-dimensional reconstruction of the connector protein from phage T3 (gp 8) from tilted view of two dimensional crystals obtained from this protein after cloning and purification.DNA sequences including gene 8 from phage T3 were cloned, into Bam Hl-Eco Rl sites down stream of lambda promotor PL, in the expression vector pNT45 under the control of cI857. E R204 (pNT89) cells were incubated at 42°C for 2h, harvested and resuspended in 20 mM Tris HC1 (pH 7.4), 7mM 2 mercaptoethanol, ImM EDTA. The cells were lysed by freezing and thawing in the presence of lysozyme (lmg/ml) and ligthly sonicated. The low speed supernatant was precipitated by ammonium sulfate (60% saturated) and dissolved in the original buffer to be subjected to gel nitration through Sepharose 6B, followed by phosphocellulose colum (Pll) and DEAE cellulose colum (DE52). Purified gp8 appeared at 0.3M NaCl and formed crystals when its concentration increased above 1.5 mg/ml.

DNA methylation in satellite repeats disorders

2019 ◽  
Vol 63 (6) ◽  
pp. 757-771 ◽  
Author(s):  
Claire Francastel ◽  
Frédérique Magdinier
Keyword(s):  
Dna Methylation ◽  
Human Genome ◽  
Repetitive Dna ◽  
Dna Sequences ◽  
Satellite Repeats ◽  
Tremendous Progress ◽  
Genes Encoding ◽  
Dna Elements ◽  
Near Future

Abstract Despite the tremendous progress made in recent years in assembling the human genome, tandemly repeated DNA elements remain poorly characterized. These sequences account for the vast majority of methylated sites in the human genome and their methylated state is necessary for this repetitive DNA to function properly and to maintain genome integrity. Furthermore, recent advances highlight the emerging role of these sequences in regulating the functions of the human genome and its variability during evolution, among individuals, or in disease susceptibility. In addition, a number of inherited rare diseases are directly linked to the alteration of some of these repetitive DNA sequences, either through changes in the organization or size of the tandem repeat arrays or through mutations in genes encoding chromatin modifiers involved in the epigenetic regulation of these elements. Although largely overlooked so far in the functional annotation of the human genome, satellite elements play key roles in its architectural and topological organization. This includes functions as boundary elements delimitating functional domains or assembly of repressive nuclear compartments, with local or distal impact on gene expression. Thus, the consideration of satellite repeats organization and their associated epigenetic landmarks, including DNA methylation (DNAme), will become unavoidable in the near future to fully decipher human phenotypes and associated diseases.

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