Journal ArticleDOI
Nucleotide sequence of the gene for human factor IX (antihemophilic factor B).
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TLDR
Two different human genomic DNA libraries were screened for the gene for blood coagulation factor IX by employing a cDNA for the human protein as a hybridization probe and found five overlapping lambda phages were identified that contained the genes for factor IX.Abstract:
Two different human genomic DNA libraries were screened for the gene for blood coagulation factor IX by employing a cDNA for the human protein as a hybridization probe. Five overlapping lambda phages were identified that contained the gene for factor IX. The complete DNA sequence of about 38 kilobases for the gene and the adjacent 5' and 3' flanking regions was established by the dideoxy chain termination and chemical degradation methods. The gene contained about 33.5 kilobases of DNA, including seven introns and eight exons within the coding and 3' noncoding regions of the gene. The eight exons code for a prepro leader sequence and 415 amino acids that make up the mature protein circulating in plasma. The intervening sequences range in size from 188 to 9473 nucleotides and contain four Alu repetitive sequences, including one in intron A and three in intron F. A fifth Alu repetitive sequence was found immediately flanking the 3' end of the gene. A 50 base pair insert in intron A was found in a clone from one of the genomic libraries but was absent in clones from the other library. Intron A as well as the 3' noncoding region of the gene also contained alternating purine-pyrimidine sequences that provide potential left-handed helical DNA or Z-DNA structures for the gene. KpnI repetitive sequences were identified in intron D and the region flanking the 5' end of the gene. The 5' flanking region also contained a 1.9-kb HindIII subfamily repeat. The seven introns in the gene for factor IX were located in essentially the same position as the seven introns in the gene for human protein C, while the first three were found in positions identical with those in the gene for human prothrombin.read more
Citations
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Journal ArticleDOI
The coagulation cascade: initiation, maintenance, and regulation
Journal ArticleDOI
The molecular basis of blood coagulation
Bruce Furie,Barbara C. Furie +1 more
TL;DR: This work focuses on the molecular basis of blood coagulation with particular attention to the biochemistry and regulation of this pathway as it relates to humans in health and disease.
Journal ArticleDOI
Evidence for gene transfer and expression of factor IX in haemophilia B patients treated with an AAV vector
Mark A. Kay,Catherine S. Manno,Catherine S. Manno,Margaret V. Ragni,Peter J. Larson,Peter J. Larson,Linda B. Couto,Alan McClelland,Bertil Glader,Amy J. Chew,Shing J Tai,Roland W. Herzog,Valder R. Arruda,Fred Johnson,Ciaran Scallan,Erik D. Skarsgard,Alan W. Flake,Alan W. Flake,Katherine A. High,Katherine A. High +19 more
TL;DR: Evidence of gene expression at low doses of vector suggests that dose calculations based on animal data may have overestimated the amount of vector required to achieve therapeutic levels in humans, and that the approach offers the possibility of converting severe haemophilia B to a milder form of the disease.
Journal ArticleDOI
Human factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts
Angelika Schnieke,Alexander Kind,W. A. Ritchie,Karen Mycock,A Scott,Marjorie Ritchie,Ian Wilmut,Alan Colman,Keith H.S. Campbell +8 more
TL;DR: Ovine primary fetal fibroblasts were cotransfected with a neomycin resistance marker gene (neo) and a human coagulation factor IX genomic construct designed for expression of the encoded protein in sheep milk and produced viable animals by nuclear transfer.
References
More filters
Journal ArticleDOI
DNA sequencing with chain-terminating inhibitors
TL;DR: A new method for determining nucleotide sequences in DNA is described, which makes use of the 2',3'-dideoxy and arabinon nucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase.
Journal ArticleDOI
Detection of specific sequences among DNA fragments separated by gel electrophoresis.
TL;DR: This paper describes a method of transferring fragments of DNA from agarose gels to cellulose nitrate filters that can be hybridized to radioactive RNA and hybrids detected by radioautography or fluorography.
Book ChapterDOI
Sequencing end-labeled DNA with base-specific chemical cleavages.
Allan M. Maxam,Walter Gilbert +1 more
TL;DR: The chapter presents techniques for producing discrete DNA fragments, end-labeling DNA, segregating end- labeled fragments, extracting DNA from gels, and the protocols for partially cleaving it at specific bases using the chemical reactions.
Journal ArticleDOI
Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I
TL;DR: Labeled DNAs (and restriction endonuclease fragments derived from them) are useful probes for detecting rare homologous sequences by in situ hybridization and reassociation kinetic analysis.
Journal ArticleDOI
Screening lambdagt recombinant clones by hybridization to single plaques in situ
WD Benton,Ronald W. Davis +1 more
TL;DR: A rapid, direct method for screening single plaques of Agt recombinant phage is described, which allows at least 10(6) clones to be screened per day and simplifies physical containment of recombinants.