POLYMORPHIC SITES IN FACTOR X GENE

1987 ◽  
Author(s):  
C Chelucci ◽  
H J Hassan ◽  
R Guerriero ◽  
A Leonardi ◽  
G Mattia ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Gene Locus ◽  
Cdna Probe ◽  
Normal Subjects ◽  
Restriction Pattern ◽  
Factor X ◽  
Congenital Deficiency ◽  
Restriction Sites ◽  
X Gene ◽  
Polymorphic Restriction

The structure of factor X gene has been analyzed by Southern blot in 5 subjects with factor X deficiency.Genomic DNA was digested with 8 different endonucleases and hybridized with a cDNA probe. The congenital deficiency observed in these patients is not apparently due to a major deletion or rearrangement. Since the gene locus is grossly intact, the disease presumably results from point mutation(s) not identified by the utilized endonucleases.Our study was also focused on the presence of polymorphic site(s) in the factor X gene locus. Analysis of 50 normal subjects allowed to identify several polymorphic restriction sites after digestion with EcoRI, Hind III and Pvu II.The restriction pattern obtained after Hind III digestion showed two bands of 7.3 and 6.0 Kb, while in two families an additional 7.6 Kb band was observed. Genomic DNA digested with EcoRI showed a 7.1 and 5.1 Kb fragments, and also a 6.6 Kb band with a 10% f requency.After DNA digestion with Pvu II 5.6, 2.7 and ∼1.0 Kb bands were observed. In three unrelated subjects we observed an additional 3.0 Kb fragment, in two other subjects a 3.5 kb band. Interestingly hybridization with a 178 bp cDNA subclone allowed to map the polymorphic sites in a the 3’ region of the gene locus.

scholarly journals Multiple polymorphic sites in factor X locus

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1353-1356 ◽  
Author(s):  
HJ Hassan ◽  
R Guerriero ◽  
C Chelucci ◽  
A Leonardi ◽  
G Mattia ◽  
...  
Keyword(s):  
Relative Frequency ◽  
Genomic Dna ◽  
Cdna Probe ◽  
Normal Subjects ◽  
Factor X ◽  
Polymorphic Region ◽  
Gene Deletions ◽  
Allelic Segregation ◽  
Normal Controls ◽  
Homozygous Subject

Abstract The structure of factor X (FX) gene was analyzed in five FX deficient pedigrees with four different variants of the disease, as well as in 50 normal subjects. Genomic DNA from the deficient patients and the normal controls was digested with 12 restriction endonucleases and hybridized with a FX cDNA probe. The results seemingly exclude gross gene deletions or rearrangements in the deficient patients. A variety of polymorphic sites (ie, EcoRI, HindIII, PstI, PvuII, TaqI) was observed within the FX locus and their relative frequency was established. Intriguingly, a highly polymorphic region for the PvuII endonuclease was identified and located approximately 3 kilobases (kb) from the last 3′ exon. These polymorphisms allowed us to analyze the allelic segregation in a FX deficient family and to identify a homozygous subject.

scholarly journals Multiple polymorphic sites in factor X locus

Blood ◽  
1988 ◽  
Vol 71 (5) ◽  
pp. 1353-1356
Author(s):  
HJ Hassan ◽  
R Guerriero ◽  
C Chelucci ◽  
A Leonardi ◽  
G Mattia ◽  
...  
Keyword(s):  
Relative Frequency ◽  
Genomic Dna ◽  
Cdna Probe ◽  
Normal Subjects ◽  
Factor X ◽  
Polymorphic Region ◽  
Gene Deletions ◽  
Allelic Segregation ◽  
Normal Controls ◽  
Homozygous Subject

The structure of factor X (FX) gene was analyzed in five FX deficient pedigrees with four different variants of the disease, as well as in 50 normal subjects. Genomic DNA from the deficient patients and the normal controls was digested with 12 restriction endonucleases and hybridized with a FX cDNA probe. The results seemingly exclude gross gene deletions or rearrangements in the deficient patients. A variety of polymorphic sites (ie, EcoRI, HindIII, PstI, PvuII, TaqI) was observed within the FX locus and their relative frequency was established. Intriguingly, a highly polymorphic region for the PvuII endonuclease was identified and located approximately 3 kilobases (kb) from the last 3′ exon. These polymorphisms allowed us to analyze the allelic segregation in a FX deficient family and to identify a homozygous subject.

One Missense Mutation in the Factor X Gene Causing Factor X Deficiency—Factor X Kanazawa

2001 ◽  
Vol 73 (3) ◽  
pp. 390-392 ◽  
Author(s):  
Eriko Morishita ◽  
Kazuo Yamaguchi ◽  
Hidesaku Asakura ◽  
Masanori Saito ◽  
Masahide Yamazaki ◽  
...  
Keyword(s):  
Missense Mutation ◽  
Factor X ◽  
Factor X Deficiency ◽  
X Gene

scholarly journals Naturally occurring variation in the restriction map of the amy region of Drosophila melanogaster.

Genetics ◽  
1988 ◽  
Vol 119 (3) ◽  
pp. 619-629
Author(s):  
C H Langley ◽  
A E Shrimpton ◽  
T Yamazaki ◽  
N Miyashita ◽  
Y Matsuo ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Restriction Site ◽  
Natural Populations ◽  
Alpha Amylase ◽  
Low Frequencies ◽  
Linkage Disequilibria ◽  
Restriction Sites ◽  
Site Variation ◽  
Transcriptional Units ◽  
Polymorphic Restriction

Abstract The restriction maps of 85 alleles of the Amy region of Drosophila melanogaster from natural populations were surveyed. A subset of these were also scored for allozyme phenotype and adult enzyme activity of alpha-amylase. Large insertions were found in 12% of the alleles in a 15-kb region surrounding the two transcriptional units of the duplicated Amy locus. The low frequencies at which each of these large insertions were found are consistent with earlier reports of variation in other loci. Four small deletions were found in the region 5' to the Amy genes. Each was also rare in the population. Restriction site variation provided an estimate of per nucleotide heterozygosity of 0.006. Several statistically significant linkage disequilibria were observed between four polymorphic restriction sites and the allozymes. Adult alpha-amylase activity was correlated with the allozymes and with the polymorphism at one restriction site close to the transcriptional units.

Restriction pattern analysis of genomic DNA of Frankia isolates

1985 ◽  
pp. 43-48
Author(s):  
Chung Sun An ◽  
W. Stuart Riggsby ◽  
Beth C. Mullin
Keyword(s):  
Genomic Dna ◽  
Pattern Analysis ◽  
Restriction Pattern

scholarly journals Cloning and characterization of platelet factor 4 cDNA derived from a human erythroleukemic cell line

Blood ◽  
1987 ◽  
Vol 69 (1) ◽  
pp. 219-223 ◽  
Author(s):  
M Poncz ◽  
S Surrey ◽  
P LaRocco ◽  
MJ Weiss ◽  
EF Rappaport ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Cell Line ◽  
Genomic Dna ◽  
Cdna Probe ◽  
Leader Sequence ◽  
Platelet Factor 4 ◽  
Coding Region ◽  
Platelet Factor ◽  
Amino Acid Homology

Abstract We report the isolation of a platelet factor 4 (PF4) cDNA clone from a lambda gt11 expression cDNA library which was derived from a human erythroleukemic (HEL) cell line. The sequence of the DNA insert includes the 3′-untranslated region, the entire amino acid coding region for the mature PF4 protein, and a 5′ region containing coding information for an additional 18 amino acids. In addition, supplemental genomic DNA sequencing shows that the full-length leader sequence is 30 amino acids long plus an initial methionine and codes for a hydrophobic signal-like sequence which is probably involved in transmembrane transport. A single species mRNA of approximately 800 nucleotides was detected on blots of HEL cell poly(A) + RNA using a labeled PF4 cDNA probe. The human PF4 leader sequence shares some DNA, but no amino acid, homology with the 15 amino acids at the N-terminus of mature bovine PF4, suggesting rapid divergence in this region of PF4 between these two species. Sequence comparison of the coding regions of mature PF4 and gamma IP-10, a protein induced in a variety of cells following treatment with gamma-interferon, shows a corrected divergence of 76%. The divergence of a common ancestor protein into PF4 and gamma IP-10 may have accompanied the development of sophisticated immune and coagulation systems in vertebrates. The availability of cDNA and genomic DNA information for these genes in other species will be useful in studying the evolution of the coagulation and immune systems.

scholarly journals Fetal hemoglobin levels in sickle cell disease and normal individuals are partially controlled by an X-linked gene located at Xp22.2

Blood ◽  
1992 ◽  
Vol 80 (3) ◽  
pp. 816-824 ◽  
Author(s):  
GJ Dover ◽  
KD Smith ◽  
YC Chang ◽  
S Purvis ◽  
A Mays ◽  
...  
Keyword(s):  
Sickle Cell ◽  
Fetal Hemoglobin ◽  
Beta Globin ◽  
Chromosome 11 ◽  
Cell Production ◽  
Flow Cytometric ◽  
Normal Individuals ◽  
Restriction Sites ◽  
F Cells ◽  
Polymorphic Restriction

Abstract Fetal hemoglobin (Hb F) production in sickle cell (SS) disease and in normal individuals varies over a 20-fold range and is under genetic control. Previous studies suggested that variant Hb F levels might be controlled by genetic loci separate from the beta-globin complex on chromosome 11. Using microscopic radial immunodiffusion and flow cytometric immunofluorescent assays to determine the percentage of F reticulocytes and F cells in SS and nonanemic individuals, we observed that F-cell levels were significantly higher in nonanemic females than males (mean +/- SD, 3.8% +/- 3.2% v 2.7% +/- 2.3%). F-cell production as determined by F reticulocyte levels in SS females was also higher than in SS males (17% +/- 10% v 13% +/- 8%). We tested the hypothesis that F-cell production in both normal and anemic SS individuals was controlled by an X-linked locus with two alleles, high (H) and low (L). Using an algorithm to determine the 99.8% confidence interval of a normal distribution in nonanemic individuals, we estimated that males and females with at least one H allele had greater than 3.3% F cells. Comparisons of male-male or female-female SS sib pairs with discordant F reticulocyte levels distinguished two phenotypes in SS males (L, less than 12%; H, greater than 12%) and three phenotypes in SS females (LL, less than 12%; HL, 12% to 24%, HH greater than 24%). Linkage analysis using polymorphic restriction sites along the X chromosome in eight SS and one AA family localized the F-cell production (FCP) locus to Xp22.2, with a maximum lod score (logarithm of odds of linkage v independent assortment) of 4.6 at a recombination fraction of 0.04.

Identification and Cloning of a Gene Locus Encoding Peptide Synthetase of Pseudomonas fluorescens by Two Sets of PCR Primers

1999 ◽  
Vol 54 (1-2) ◽  
pp. 105-109 ◽  
Author(s):  
Narayanan Rajendran
Keyword(s):  
Bacillus Subtilis ◽  
Pseudomonas Fluorescens ◽  
Genomic Dna ◽  
Gene Locus ◽  
Genetic Locus ◽  
Pcr Primers ◽  
Chromosomal Locus ◽  
Sequence Alignments ◽  
Peptide Synthetases ◽  
Peptide Synthetase

A chromosomal locus encoding biosynthetic genes for a putative peptide synthetase of Pseudomonas fluorescens was identified and cloned. To achieve this, two sets of degenerated oligonucleotide primers KAGGA:SGTTG and TGD :LGG were used in PCR. These primers were selected based on highly conserved units of known peptide synthetases involved in adenylation and thiolation regions of Bacillus subtilis. The discrete amplified bands from PCR ca. 300 bp for KAGGA:SGTTG and ca. 500 bp for TGD:LGG proved to be integral part of the genomic DNA of P. fluorescens were cloned and sequenced. Sequence alignments of both fragments confirmed the putative peptide synthetase genes in P. fluorescens. The present study describes the identification and cloning of peptide synthetase genes of P. fluorescens, which can be used to identify a genetic locus encoding peptide synthetase in other microbial species

scholarly journals Molecular defect in coagulation factor XFriuli results from a substitution of serine for proline at position 343

Blood ◽  
1991 ◽  
Vol 77 (2) ◽  
pp. 317-323 ◽  
Author(s):  
HL James ◽  
A Girolami ◽  
DS Fair
Keyword(s):  
Point Mutation ◽  
Serine Proteases ◽  
Direct Sequencing ◽  
Coagulation Factor ◽  
Amino Acid Position ◽  
Molecular Defect ◽  
Nucleotide Position ◽  
Factor X ◽  
Gene Coding ◽  
X Gene

Abstract Our previous findings suggested that coagulation factor XFriuli could be functionally defective owing to a point mutation in the portion of the factor X gene coding for the fully activated heavy chain. To verify the existence of this postulated change, we analyzed all eight exons of the normal and Friuli factor X gene. Each exon was amplified from genomic DNA using the polymerase chain reaction and cloned into the plasmid pUC19. The amplified DNA inserts were subjected to direct sequencing by the dideoxy chain termination method with forward and reverse oligonucleotide sequencing primers. A point mutation (C to T transition at nucleotide position 19,297) that results in coding for serine (TCC) in place of proline (CCC) at amino acid position 343 was found. This substitution involves a highly conserved proline residue oriented spatially close to both the cleavage site of the zymogen and the active site of the enzyme and explains the previous observations of discrete biochemical and functional differences between factor XFriuli and normal factor X. The mutation abolished an HgiCI restriction site present in the normal factor X gene, and this change constitutes the basis for a convenient method for screening individuals carrying this molecular defect. Proline343 is in conserved region 5 of the serine protease superfamily to which factor X belongs and is part of a 14- residue L*****P******C motif that occurs in at least 16 other enzymes. Computer analysis suggests that the motif may be an essential aspect of conformational features important to functional properties of factor X as well as other serine proteases.

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