scholarly journals Chemiluminescent hybridization-ligation assays for delta F508 and delta I507 cystic fibrosis mutations

1996 ◽  
Vol 42 (1) ◽  
pp. 14-18 ◽  
Author(s):  
R A Martinelli ◽  
J C Arruda ◽  
P Dwivedi
Keyword(s):  
Cystic Fibrosis ◽  
Dna Sequences ◽  
Gel Electrophoresis ◽  
Dna Ligase ◽  
Single Nucleotide ◽  
Dna Oligomers ◽  
Synthetic Dna ◽  
Human Dna ◽  
Convenient Means ◽  
Acridinium Ester

Abstract Chemiluminescent hybridization-ligation assays were devised to detect the delta F508 and delta I507 cystic fibrosis mutations in samples of human DNA that had been amplified by PCR. Two synthetic DNA oligomers were used in each assay. One of the oligomers was labeled with an acridinium ester and the other was immobilized on paramagnetic particles. The oligomers were hybridized to the samples and the target sequences discriminated by ligation with T4 or a thermostable DNA ligase. The performance of the assay was evaluated in a blind study of 30 samples. There was complete correspondence between the assignments based on the chemiluminescent assay and those made previously by gel electrophoresis, with one exception. The assignment of this discrepant sample by the chemiluminescent assay as a delta I507/normal heterozygote rather than a delta F508/normal heterozygote was confirmed by sequencing. The chemiluminescent hybridization-ligation assay provides a rapid and convenient means of discriminating DNA sequences differing by a single nucleotide.

scholarly journals Heteroalleles in Common Wheat: Multiple Differences between Allelic Variants of the Gli-B1 Locus

2021 ◽  
Vol 22 (4) ◽  
pp. 1832
Author(s):  
Eugene Metakovsky ◽  
Laura Pascual ◽  
Patrizia Vaccino ◽  
Viktor Melnik ◽  
Marta Rodriguez-Quijano ◽  
...  
Keyword(s):  
Common Wheat ◽  
Dna Sequences ◽  
Fragment Length Polymorphism ◽  
Snp Markers ◽  
Group Iv ◽  
Nucleotide Polymorphisms ◽  
High Genetic Diversity ◽  
Single Nucleotide ◽  
Allelic Variants ◽  
B Genome

The Gli-B1-encoded γ-gliadins and non-coding γ-gliadin DNA sequences for 15 different alleles of common wheat have been compared using seven tests: electrophoretic mobility (EM) and molecular weight (MW) of the encoded major γ-gliadin, restriction fragment length polymorphism patterns (RFLPs) (three different markers), Gli-B1-γ-gliadin-pseudogene known SNP markers (Single nucleotide polymorphisms) and sequencing the pseudogene GAG56B. It was discovered that encoded γ-gliadins, with contrasting EM, had similar MWs. However, seven allelic variants (designated from I to VII) differed among them in the other six tests: I (alleles Gli-B1i, k, m, o), II (Gli-B1n, q, s), III (Gli-B1b), IV (Gli-B1e, f, g), V (Gli-B1h), VI (Gli-B1d) and VII (Gli-B1a). Allele Gli-B1c (variant VIII) was identical to the alleles from group IV in four of the tests. Some tests might show a fine difference between alleles belonging to the same variant. Our results attest in favor of the independent origin of at least seven variants at the Gli-B1 locus that might originate from deeply diverged genotypes of the donor(s) of the B genome in hexaploid wheat and therefore might be called “heteroallelic”. The donor’s particularities at the Gli-B1 locus might be conserved since that time and decisively contribute to the current high genetic diversity of common wheat.

Human DNA sequences homologous to a protein coding region conserved between homeotic genes of Drosophila

Cell ◽  
1984 ◽  
Vol 38 (3) ◽  
pp. 667-673 ◽  
Author(s):  
Michael Levine ◽  
Gerald M. Rubin ◽  
Robert Tjian
Keyword(s):  
Dna Sequences ◽  
Homeotic Genes ◽  
Coding Region ◽  
Protein Coding ◽  
Human Dna

scholarly journals Human DNA Ligase III Recognizes DNA Ends by Dynamic Switching between Two DNA-Bound States

Biochemistry ◽  
2010 ◽  
Vol 49 (29) ◽  
pp. 6165-6176 ◽  
Author(s):  
Elizabeth Cotner-Gohara ◽  
In-Kwon Kim ◽  
Michal Hammel ◽  
John A. Tainer ◽  
Alan E. Tomkinson ◽  
...  
Keyword(s):  
Bound States ◽  
Dna Ligase ◽  
Dynamic Switching ◽  
Human Dna ◽  
Dna Ends ◽  
Dna Ligase Iii

scholarly journals Molecular cloning and chromosomal localization of DNA sequences associated with a human DNA repair gene.

1985 ◽  
Vol 5 (2) ◽  
pp. 398-405 ◽  
Author(s):  
J S Rubin ◽  
V R Prideaux ◽  
H F Willard ◽  
A M Dulhanty ◽  
G F Whitmore ◽  
...  
Keyword(s):  
Dna Repair ◽  
Dna Sequences ◽  
Chromosomal Localization ◽  
Excision Repair ◽  
Repair Process ◽  
Human Cells ◽  
Gene Products ◽  
Repair Gene ◽  
Human Dna ◽  
Dna Repair Gene

The genes and gene products involved in the mammalian DNA repair processes have yet to be identified. Toward this end we made use of a number of DNA repair-proficient transformants that were generated after transfection of DNA from repair-proficient human cells into a mutant hamster line that is defective in the initial incision step of the excision repair process. In this report, biochemical evidence is presented that demonstrates that these transformants are repair proficient. In addition, we describe the molecular identification and cloning of unique DNA sequences closely associated with the transfected human DNA repair gene and demonstrate the presence of homologous DNA sequences in human cells and in the repair-proficient DNA transformants. The chromosomal location of these sequences was determined by using a panel of rodent-human somatic cell hybrids. Both unique DNA sequences were found to be on human chromosome 19.

Stm Studies at Electrochemically Controlled Interfaces

1994 ◽  
Vol 332 ◽  
Author(s):  
S.M. Lindsay ◽  
J. Pan ◽  
T.W. Jing
Keyword(s):  
Electron Tunneling ◽  
Point Contact ◽  
Localized States ◽  
Scanning Tunneling ◽  
Zero Field ◽  
Tunneling Microscopy ◽  
Dna Oligomers ◽  
Synthetic Dna ◽  
Quantum Point

ABSTRACTWe use electrochemical methods to control the adsorption of molecules onto an electrode for imaging in-situ by scanning tunneling microscopy. Measurements of the barrier for electron tunneling show that the mechanism of electron transfer differs from vacuum tunneling. Barriers depend upon the direction of electron tunneling, indicating the presence of permanently aligned dipoles in the tunnel gap. We attribute a sharp dip in the barrier near zero field to induced polarization. We propose a ‘tunneling’ process consisting of two parts: One is delocalization of quantum-coherent states in parts of the molecular adlayer that hybridize strongly (interaction ≥ kT) with Bloch states in the metal. This gives rise to a quantum-point-contact conductance, Gc ≤ 2e2/h at a height zo. The other part comes from the exponential decay of the tails of localized states, G = Gc exp{−2K(z − z0)}. Because measured decay lengths, (2K‘)−1, are small (≈ 1 Å), STM contrast is dominated by the contour along which G[z0 (x,y)] = Gc. Measured changes in z0 are used to calculate images which are in reasonable agreement with observations. We illustrate this with images of synthetic DNA oligomers.

Molecular cloning and characterization of an activated human c-raf-1 gene

1987 ◽  
Vol 7 (5) ◽  
pp. 1776-1781
Author(s):  
M Fukui ◽  
T Yamamoto ◽  
S Kawai ◽  
F Mitsunobu ◽  
K Toyoshima
Keyword(s):  
Dna Sequences ◽  
Promoter Sequence ◽  
Small Population ◽  
Dna Transfection ◽  
Amino Terminal ◽  
Human Dna ◽  
Nih 3T3 ◽  
Amino Terminal Sequence ◽  
Tumor Dna

Results of previous studies have shown that a raf-related transforming DNA sequence is present in NIH 3T3 transformants that are derived from GL-5-JCK human glioblastoma DNA transfection. The transforming DNA was molecularly cloned by using cosmid vector pJB8 to determine its structure and origin. Analyses of selected clones revealed that the transforming DNA consisted of three portions of human DNA sequences, with the 3' half of the c-raf-1 gene as its middle portion. This raf region was about 20 kilobases long and contained exons 8 to 17 and the poly(A) addition site. RNA blot analysis showed that the raf-related transforming DNA was transcribed into 5.3-, 4.8-, and 2.5-kilobase mRNAs; the 2.5-kilobase transcript was thought to be the major transcript. Immunoprecipitation analyses revealed that a 44-kilodalton raf-related protein was specifically expressed in the NIH 3T3 transformants. The raf-related transforming DNA was considered to be activated when its amino-terminal sequence was truncated and the DNA was coupled with a foreign promoter sequence. On hybridization analysis of the original GL-5-JCK glioblastoma DNA, no rearrangement of c-raf-1 was detectable in the tumor DNA. The rearrangement of c-raf-1 may have occurred during transfection or may have been present in a small population of the original tumor cells as a result of tumor progression.

Oncogenes in Laryngeal Cancer: Serial Passage of Transformed Cellular DNA

1985 ◽  
Vol 93 (3) ◽  
pp. 346-350 ◽  
Author(s):  
William H. Friedman ◽  
Barry N. Rosenblum ◽  
Paul Loewenstein ◽  
Helen Thornton ◽  
George Katsantonis ◽  
...  
Keyword(s):  
Laryngeal Cancer ◽  
Dna Sequences ◽  
Squamous Cell Cancer ◽  
Cell Cancer ◽  
Serial Passage ◽  
3T3 Cells ◽  
Human Dna ◽  
Cellular Dna ◽  
Nih 3T3 ◽  
Narrow Bands

DNA originally extracted from squamous cell cancer of the larynx has been serially passaged through transformed populations of NIH/3T3 mouse fibroblasts. The transformed foci were then harvested, cloned to volume, and incubated with a fresh population of NIH/3T3 cells in a second passage. Transforming efficiencies were enhanced by serial passage. In addition, Southern Blot analysis of the transformed foci revealed hybridization between transformant DNA and human probe DNA from the Alu family of conserved human DNA sequences. In the first passage this hybridization took the form of diffuse homology throughout the entire molecular weight distribution. The second-passage DNA showed “narrow bands” indicating the possibility that an oncogene has been identified in laryngeal cancer and that serial passage has eliminated contaminating human sequences. Repetitive transfection in third- and fourth-passage studies is now being completed.

scholarly journals Novel Endogenous Retrovirus in Rabbits Previously Reported as Human Retrovirus 5

2002 ◽  
Vol 76 (14) ◽  
pp. 7094-7102 ◽  
Author(s):  
David J. Griffiths ◽  
Cécile Voisset ◽  
Patrick J. W. Venables ◽  
Robin A. Weiss
Keyword(s):  
Sequence Analysis ◽  
Dna Sequences ◽  
Inflammatory Diseases ◽  
Endogenous Retrovirus ◽  
European Rabbit ◽  
Pcr Analysis ◽  
Northern Hybridization ◽  
Human Dna ◽  
Integration Sites ◽  
Human Retrovirus

ABSTRACT Human retrovirus 5 (HRV-5) represented a fragment of a novel retrovirus sequence identified in human RNA and DNA preparations. In this study, the genome of HRV-5 was cloned and sequenced and integration sites were analyzed. Using PCR and Southern hybridization, we showed that HRV-5 is not integrated into human DNA. A survey of other species revealed that HRV-5 is present in the genomic DNA of the European rabbit (Oryctolagus cuniculus) and belongs to an endogenous retrovirus family found in rabbits. The presence of rabbit sequences flanking HRV-5 proviruses in human DNA extracts suggested that rabbit DNA was present in our human extracts, and this was confirmed by PCR analysis that revealed the presence of rabbit mitochondrial DNA sequences in four of five human DNA preparations tested. The origin of the rabbit DNA and HRV-5 in human DNA preparations remains unclear, but laboratory contamination cannot explain the preferential detection of HRV-5 in inflammatory diseases and lymphomas reported previously. This is the first description of a retrovirus genome in rabbits, and sequence analysis shows that it is related to but distinct from A-type retroelements of mice and other rodents. The species distribution of HRV-5 is restricted to rabbits; other species, including other members of the order Lagomorpha, do not contain this sequence. Analysis of HRV-5 expression by Northern hybridization and reverse transcriptase PCR indicates that the virus is transcribed at a low level in many rabbit tissues. In light of these findings we propose that the sequence previously designated HRV-5 should now be denoted RERV-H (for rabbit endogenous retrovirus H).

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