共查询到20条相似文献,搜索用时 31 毫秒
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Chih-Ming Chou Wen-chang Lin Jiann-Horng Leu Ter-Lee Su Chen-Kung Chou Dr. Chang-Jen Huang 《Journal of biomedical science》1998,5(2):127-134
The round-spotted pufferfishTetraodon fluviatilis has a genome size of 380 Mb which is slightly smaller than that of another pufferfish,Fugu rubripes rubripes (Fugu). Due to their compact genome and small introns, both pufferfishes have been proposed as model organisms for genome studies. In this study, we have used genomic DNA as template to perform PCR to screen for protein kinase (pk) genes. Forty-oneT. fluviatilis pk genes encoding 7 receptor tyrosine kinases, 14 nonreceptor tyrosine kinases, 16 serine/threonine kinases, 1 dual kinase and 3 novel kinases have been identified. The success of this approach depends on the size and location of the introns. Most of the identifiedpk gene fragments contain introns, ranging from 71 to 300 bp, with an average of 120 bp. It is noteworthy that the intron/exon boundaries of certain genes which belong to the same family are identical. We also analyzed by specific RT-PCR primers the expression profile of those 3 novel genes as well as some selectedpk genes in a variety of tissues. We found thaterbB3,pku , mrk, CaMK I,CaMKII, and two novel kinase genes (133 and 3–26) are expressed in all tissues examined. However, the novel clone 146 is strongly expressed in the brain and weakly in the intestine, kidney and heart. 相似文献
3.
Toshio Itoh T. Tanaka Ryozo Nagai Kenjiro Kikuchi Satoshi Ogawa Shintaro Okada Shiro Yamagata Katsusuke Yano Yoshio Yazaki Yusuke Nakamura 《Human genetics》1998,103(3):290-294
To elucidate the role of the KVLQT1 gene in the pathogenesis of long QT syndrome (LQTS), we have established a screening system for detecting KVLQT1 mutations by the polymerase chain reaction-single strand conformation polymorphism technique (PCR-SSCP). We first determined
exon/intron boundaries and flanking intronic sequences, and found that the KVLQT1 gene consists of 17 coding exons. Subsequently, we synthesized oligonucleotide primers to cover the coding region and the
flanking intronic sequences, and searched for mutations in 31 Japanese LQTS families. When genomic DNA from each proband was
examined by PCR-SSCP followed by direct DNA sequencing, mutations were detected in five families; two independent families
carried the same mutation and three of the four were novel. Each mutation was present in affected relatives of the respective
proband. This work will enable us to search more thoroughly for LQTS mutations associated with KVLQT1, and eventually will help us in finding genotype/phenotype relationships.
Received: 20 March 1998 / Accepted: 30 April 1998 相似文献
4.
Toshio Itoh T. Tanaka Ryozo Nagai Tetsuro Kamiya Toshitami Sawayama Toshio Nakayama Hitonobu Tomoike Harumizu Sakurada Yoshio Yazaki Yusuke Nakamura 《Human genetics》1998,102(4):435-439
Familial long QT syndrome (LQTS) is characterized by prolonged ventricular repolarization. Clinical symptoms include recurrent
syncopal attacks, and sudden death may occur as a result of ventricular tachyarrhythmias. Three genes responsible for this
syndrome (KVLQT1, HERG, and SCN5A) have been identified so far, and mutations have been reported on the basis of partially characterized genomic organization.
To optimize the search for HERG mutations, we have determined the genomic structure of HERG and investigated mutations in LQTS families. Human genomic clones containing the HERG gene were isolated from a human genomic library by using reverse-transcribed polymerase chain reaction (RT-PCR) products
from this gene as probes. We determined exon/intron boundaries and flanking intronic sequences by using primers synthesized
on the basis of the HERG cDNA sequence available in the DNA database. HERG was shown to consist of 15 exons spanning approximately 19 kb on chromosome 7q35. Subsequently, we synthesized oligonucleotide
primers to cover the entire coding region and searched for mutations in 36 Japanese LQTS families. When genomic DNA from each
proband was examined by the PCR/single-strand conformation polymorphism technique followed by direct DNA sequencing, five
novel mutations were detected. Each mutation was present in affected relatives of the respective proband. This work should
increase the efficiency of screening mutations associated with HERG.
Received: 4 November 1997 / Accepted: 5 January 1998 相似文献
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C. Settasatian S.A. Whitmore J. Crawford R.L. Bilton A.-M. Cleton-Jansen G.R. Sutherland D.F. Callen 《Human genetics》1999,105(1-2):139-144
SPG7 is a newly identified gene involved in an autosomal recessive form of hereditary spastic paraplegia (HSP), a genetically
heterogeneous group of neurodegenerative disorders. This gene encodes a protein characterized as a nuclear-encoded mitochondrial
metalloprotease. The present report describes the genomic structure of the SPG7 gene. It is organized into 17 exons ranging from 78 to 242 bp and spans approximately 52 kb within three overlapping cosmids.
The exon/intron boundaries and all splice junctions are consistent with the published consensus sequences for donor and acceptor
sites. The provided genomic structure of SPG7 should facilitate the screening for mutations in this gene in patients with HSP and other related mitochondrial disease syndromes. SPG7 has been mapped to chromosome 16q24.3, a region of frequent loss of heterozygosity (LOH) seen in sporadic breast and prostate
cancer. We have performed single-strand conformation polymorphism analysis of ten exons of this gene in a number of sporadic
breast cancer samples showing LOH at 16q24.3. No mutations were detected; only single nucleotide polymorphisms were observed
in exon 11, intron 7, intron 10 and intron 12. An expression analysis study has revealed the differential expression of SPG7 mRNA in various tissues and at different developmental stages.
Electronic Publication 相似文献
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Stöhr H Klein J Gehrig A Koehler MR Jurklies B Kellner U Leo-Kottler B Schmid M Weber BH 《Human genetics》1999,104(1):99-105
The family of diacylglycerol kinases (DAGKs) is known to play an important role in signal transduction linked to phospholipid
turnover. In the fruitfly Drosophila melanogaster, a human DAGK ortholog, DGK2, was shown to underlie the phenotype of the visual mutant retinal degeneration A (rdgA). Previously, the gene encoding a novel member of the human DAGK family, termed DAGK3, was cloned and demonstrated to be abundantly expressed in the human retina. Based on these findings we reasoned that DAGK3 might be an excellent candidate gene for a human eye disease. In the present study, we report the genomic organization of
the human DAGK3 gene, which spans over 30 kb of genomic DNA interrupted by 23 introns. In addition, we have mapped the gene locus by fluorescence
in situ hybridization to 3q27–28, overlapping the chromosomal region known to contain the gene underlying dominant optic atrophy
(OPA1), the most common form of hereditary atrophy of the optic nerve. Mutational analysis of the entire coding region of
DAGK3 in 19 unrelated German OPA1 patients has not revealed any disease-causing mutations, therefore excluding DAGK3 as a major cause underlying OPA1.
Received: 24 August 1998 / Accepted: 13 October 1998 相似文献
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DNA sequence, structure, and tyrosine kinase activity of the Drosophila melanogaster Abelson proto-oncogene homolog. 总被引:12,自引:4,他引:8
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M J Henkemeyer R L Bennett F B Gertler F M Hoffmann 《Molecular and cellular biology》1988,8(2):843-853
We report our molecular characterization of the Drosophila melanogaster Abelson gene (abl), a gene in which recessive loss-of-function mutations result in lethality at the pupal stage of development. This essential gene consists of 10 exons extending over 26 kilobase pairs of genomic DNA. The DNA sequence encodes a protein of 1,520 amino acids with strong sequence similarity to the human c-abl proto-oncogene beginning in the type lb 5' exon and extending through the region essential for tyrosine kinase activity. When the tyrosine kinase homologous region was expressed in Escherichia coli, phosphorylation of proteins on tyrosine residues was observed with an antiphosphotyrosine antibody. These results show that the abl gene is highly conserved through evolution and encodes a functional tyrosine protein kinase required for Drosophila development. 相似文献
11.
Baojin Wu Yixiang Shao Bing Chen Chun Liu Zhengfeng Xue Peilin Wu Houda Li 《Cell biochemistry and biophysics》2010,58(3):129-135
Mutations in T-box genes are associated with numerous disease states in humans. The objective of this paper was to characterize
the T
shao
, a specific T-box mutation, in mice. T
shao
, a short-tailed mutant mouse strain in a B6 background, was obtained by ethylnitrosourea mutagenesis. Microsatellite genomic
scans mapped the location of the mutation. RT–PCR was used to amplify the identified region and the product was sequenced.
DNA of the region was sequenced and scanned for mutations. Tails of T
shao
mice were mostly curly with tail length ranging from less than 1 cm (tail bud) to half of the normal length. T
shao
presented single dominance gene inheritance, and homozygous mutant mice died approximately at E10. Scans of the F2 generation
mapped the mutant gene to chromosome 17, near D17Mit143. The Brachyury (T) gene was identified as a potential candidate gene in this location. To confirm this, RT–PCR was performed on RNA from intercrossed
8.5-day embryos, and products were sequenced. A 67-nucleotide deletion in exon 2 of the mutant T gene was identified. Further sequencing of the genomic DNA from this region identified a T to A transversion at the 67th
nucleotide of exon 2. The T
shao
mutation is a result of a deletion in exon 2 causing the early termination and loss of function of protein encoded by the
T gene, manifesting as a short tail phenotype. 相似文献
12.
Shimbata T Nakamura T Vrinten P Saito M Yonemaru J Seto Y Yasuda H 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2005,111(6):1072-1079
Wheat (Triticum aestivum L.) starch synthase II, which is also known as starch granule protein 1 (SGP-1), plays a major role in endosperm starch synthesis. The three SGP-1 proteins, SGP-A1, B1 and D1, are produced by three homoeologous SSII genes, wSSII-A, B, and D. Lines carrying null alleles for each SGP-1 protein have previously been identified. In this report, the mutations occurring in each wSSII gene were characterized, and PCR-based DNA markers capable of detecting the mutations were developed. In the null wSSII-A allele, a 289 bp deletion accompanied by 8 bp of filler DNA was present near the initiation codon. A 175 bp insertion occurred in exon 8 of the null wSSII-B allele. The insertion represented a recently discovered miniature inverted-repeat transposable element (MITE) named Hikkoshi that was first found in a wheat waxy gene. A 63 bp deletion was found at the region surrounding the junction of the fifth exon and intron of the null wSSII-D allele. Based on this information, we designed primer sets to enable us to conduct allele-specific amplifications for each locus. The applicability of these primer sets for breeding programs was demonstrated by reconstructing a line lacking all three SGP-1 proteins using marker-assisted selection. These markers will also be useful in breeding programs aimed at obtaining partial mutants missing one or two SGP-1 proteins. 相似文献
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14.
A novel nonreceptor tyrosine kinase, Srm: cloning and targeted disruption. 总被引:2,自引:1,他引:1
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N Kohmura T Yagi Y Tomooka M Oyanagi R Kominami N Takeda J Chiba Y Ikawa S Aizawa 《Molecular and cellular biology》1994,14(10):6915-6925
15.
Ming Mai Haojie Huang Christopher Reed Chiping Qian Justin S. Smith Benjamin Alderete Robert Jenkins David I. Smith Wanguo Liu 《Genomics》1998,51(3):359
p73, a protein having substantial structural and functional similarity to p53, has recently been identified and demonstrated to be a potential tumor suppressor. Its location on human chromosome 1p36.33 implicates p73 as a candidate for neuroblastoma. Like neuroblastoma, oligodendrogliomas also show a high frequency of deletions in chromosome 1p36.3. To determine whetherp73is a potential tumor suppressor gene involved in the development of oligodendrogliomas, we performed mutation analysis ofp73in oligodendrogliomas with chromosome 1 p-arm deletions. We first determined the genomic organization and the intron–exon boundary sequences of thep73gene by long PCR, vectorette PCR, and Southern hybridization. This gene spans about 65 kb with a large first intron. Primer pairs for the amplification of each of the 13 p73 encoding exons were designed in corresponding introns. The amplicons were then analyzed using the denaturing high-performance liquid chromatography system for mutations in thep73gene. Twenty oligodendroglioma samples with 1p36.3 deletions were screened, but no mutations were detected except for several polymorphisms. It is thus clear thatp73is not a candidate gene for oligodendroglioma despite its location in the frequently deleted 1p36.3 region. 相似文献
16.
Evelin Schröck Priscilla Badger Denise Larson Michael Erdos Anthony Wynshaw-Boris Thomas Ried Lawrence Brody 《Human genetics》1996,97(2):256-259
The recently cloned human breast and ovarian cancer suseptibility gene,BRCA1, is located on human chromosome 17q21. We have isolated murine genomic clones containingBrca1 as a first step in generating a mouse model for the loss ofBRCA1 function. A mouse genomic library was screened using probes corresponding to exon 11 of the humanBRCA1 gene. Two overlapping mouse clones were identified that hybridized to humanBRCA1 exons 9–12. Sequence analysis of 1.4 kb of the region of these clones corresponding to part of human exon 11 revealed 72% nucleic acid identity but only 50% amino acid identity with the human gene. The longest of the mouseBrca1 genomic clones maps to chromosome 11D, as determined by two-color fluorescence in situ hybridization. The synteny to human chromosome 17 was confirmed by cohybridization with the mouse probe for the NF1-gene. This comparative study confirms that the relative location of theBRCA1 gene has been conserved between mice and humans. 相似文献
17.
I. Zito D.L. Thiselton M.B. Gorin J.T. Stout C. Plant A.C. Bird S.S. Bhattacharya A.J. Hardcastle 《Human genetics》1999,105(1-2):57-62
The X-linked form of retinitis pigmentosa (XLRP) is a severe disease of the retina, characterised by night blindness and
visual field constriction in a degenerative process, culminating with complete loss of sight within the third decade of life.
Genetic mapping studies have identified two major loci for XLRP: RP3 (70%–75% of XLRP) and RP2 (20%–25% of XLRP). The RPGR (retinitis pigmentosa GTPase regulator) gene has been cloned within the RP3 genomic interval and it has been shown that 10%–20%
of XLRP families have mutations in this gene. Here, we describe a single-strand conformational polymorphism-based mutation
screening of RPGR in a pool of 29 XLRP families for which the disease segregates with the RP3 locus, in order to investigate the proportion of RP3 families with RPGR mutations and to relate the results to previous reports. Five different new mutations have been identified: two splice site
mutations for exon 1 and three frameshift mutations in exons 7, 10 and 11. The percentage of RPGR mutations identified is 17% (5/29) in our genetically well-defined population. This figure is comparable to the percentage
of RP2 gene mutations that we have detected in our entire XLRP patient pool (10%–15%). A correlation of RPGR mutations with phenotype in the families described in this study and the biochemical characterisation of reported mutations
may provide insights into the function of the protein.
Electronic Publication 相似文献
18.
Alfred L. George Jr. Geeta S. Iyer Robert Kleinfield Roland G. Kallen Robert L. Barchi 《Genomics》1993,15(3)
Voltage-dependent sodium channels are essential for normal membrane excitability and contractility in adult skeletal muscle. The gene encoding the principal sodium channel α-subunit isoform in human skeletal muscle (SCN4A) has recently been shown to harbor point mutations in certain hereditary forms of periodic paralysis. We have carried out an analysis of the detailed structure of this gene including delineation of intron-exon boundaries by genomic DNA cloning and sequence analysis. The complete coding region of SCN4A is found in 32.5 kb of genomic DNA and consists of 24 exons (54 to > 2.2 kb) and 23 introns (97 bp-4.85 kb). The exon organization of the gene shows no relationship to the predicted functional domains of the channel protein and splice junctions interrupt many of the transmembrane segments. The genomic organization of sodium channels may have been partially conserved during evolution as evidenced by the observation that 10 of the 24 splice junctions in SCN4A are positioned in homologous locations in a putative sodium channel gene in Drosophila (para). The information presented here should be extremely useful both for further identifying sodium channel mutations and for gaining a better understanding of sodium channel evolution. 相似文献
19.
Isolation of chicken cellular DNA sequences with homology to the region of viral oncogenes that encodes the tyrosine kinase domain. 总被引:8,自引:3,他引:5
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D A Foster J B Levy G Q Daley M C Simon H Hanafusa 《Molecular and cellular biology》1986,6(1):325-331
A library of chicken genomic DNA was screened for sequences that could hybridize to a cloned DNA fragment containing the transforming gene (v-fps) of Fujinami sarcoma virus. In addition to c-fps, two unique chicken cellular DNA sequences were isolated that hybridized weakly to v-fps. These sequences hybridized with many other viral oncogenes encoding tyrosine kinases. Sequence analysis of the region where homology was detected revealed a region that is highly conserved among the tyrosine kinases both at the nucleotide and amino acid levels. Although we were unable to detect expression of either chicken cellular DNA sequence in a variety of avian tissues, the data suggest the existence of additional members of the tyrosine kinase gene family. Screening genomic libraries for sequences that hybridize weakly to functional regions of other genes may prove useful for the isolation and characterization of additional members of other gene families. 相似文献
20.
The most telomeric class I region of the MHC in rat and mouse is the M region, which contains about 20 class I genes or gene fragments. The central part carries three class I genes—M4, M5, and M6—which are orthologous between the two species. M4 and M6 are pseudogenes in the mouse but transcribed, intact genes in the rat. To analyze the pseudogene status for the mouse genes in more detail, we have sequenced the respective exons in multiple representative haplotypes. The stop codons are conserved in all mouse strains analyzed, and, consistent with the pseudogene status, all strains show additional insertions and deletions, taking the genes further away from functionality. Thus, M4 and M6 indeed have a split status. They are silent in the mouse but intact in the closely related rodent, the rat.GenBank accession numbers: AF057065 to AF057072 (exon 3 of H2-M4 of reported mouse strains), AF057976 to AF057985 (exon 3 of RT1.M4 of reported rat strains), AF058923 and AF058924 (exon 2 of RT1.M4 of strains PVG and BN), AY286080 to AY286092 (exon 4 of H2-M6 of reported mouse stains), and AY303772 (full-length genomic sequence of RT1.M6-1l) 相似文献