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1.
14个染色体区特异性探针池的构建 总被引:9,自引:2,他引:7
本文运用人类染色体显微切割和PCR技术,成功地构建了14个染色体区带专特性探针池,并通过染色体原位杂交证明它们均分别来源于相应的被切割的染色体区带。 相似文献
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应用端粒区带涂染探针检测染色体微小结构重排 总被引:1,自引:0,他引:1
为了评估染色体端粒区带涂染探针在遗传诊断的应用价值,应用显微切割获得的11q、12q和22q等3个染色体端粒区涂染探针(11q23.3→qter,12q24.1→qter,22q13.1→qter),通过荧光原位杂交技术分析两个疑有染色体末端微小易位的习惯性流产病例。结果显示,病例1和病例2分别为t(11;12)和t(11;22)长臂末端间的微小易位,结合G显带技术确定断裂位点位于11q23.3、12q24.1、22q13.1。结果表明特异性染色体端粒区带探针可以确诊染色体末端区域的微小结构异常,可作为一种检出隐匿易位携带者并确定断裂位点的方法。 相似文献
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本文报道了从显微切割的人染色体区带直接分离区带专一性表达序列的方法和结果。以区带探针池为基础构建了单拷贝DNA分子库,并以该库和人骨髓细胞cDNA分子库为主要研究材料,从5×10~5个DNA克隆中筛选到68个初级阳性克隆,复筛得到了32个次级阳性克隆,分别命名为cFD14-1~32。再经14q24.3 DNA、17q11-12 DNA和人基因组总DNA作dot blot DNA杂交验证,最终得到24个14q24.3区带专一性表达顺序。测定了其中13个片段的部分序列,在NCBI数据库查新,均为新的cDNA片段,并与某些已知基因有一定的同源性。其中cFD14-1的初步表达谱分析提示了本实验所得cDNA片段均有可能用来进一步筛选位于14q24.3区带内的尚未克隆的基因。 相似文献
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染色体微切割和微克隆已成为复杂基因组研究的有效途径,但是操作过程中的核外DNA的污染一直是令人担心的问题.通过研究植物染色体微切割(微分离)和微切割的染色体DNA 扩增过程中细胞质DNA的污染问题,表明目前常用的植物染色体微切割过程中,细胞质DNA的污染几乎难以避免,并提出了一个改进的降低细胞质DNA污染的方法,对如何控制细胞质DNA的污染进行了详细的讨论. 相似文献
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染色体微切割和微克隆已成为复杂基因组研究的有效途径。但是操作过程中的核外DNA的污染一直是令人担心的问题,通过研究植物染色体微切割(微分离)和微切割的染色体DNA扩增过程中细胞质DNA的污染问题,表明目前常用的植物染色体微切割过程中,细胞质DNA的污染几乎难以避免,并提出了一个改进的降低细胞质DNA污染的方法。对如何控制细胞质DNA的污染进行了详细的讨论。 相似文献
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Weise A Rittinger O Starke H Ziegler M Claussen U Liehr T 《Cytogenetic and genome research》2003,103(1-2):14-16
We report on a moderately mentally retarded 12-year-old boy of short stature showing the most complex chromosomal rearrangement (CCR) within a single chromosome ever described. A de novo derivative chromosome 21 was recognized in GTG-banding shortly after birth. However, the nature of the rearrangement remained obscure up to the application of the chromosome 21-specific centromere-near multicolor-FISH (subcenM-FISH) probe set and of six selected locus-specific probes along chromosome 21. An unbalanced 9-break-event was uncovered with breakpoints in 21p13, 21p13-->12, 21q11.2, 21q21.1, 21q22.11, 21q22.11, 21q22.12, 21q22.22 and 21q22.3. A deletion of 21q22.11 was detected by application of the BAC probe bk249H10. The karyotype can be described as 46,XY,der(21)(:p13-->p1213::q22.3-->q22.22:: q11.2-->p1213::q11.2-->q21.1::q22.11-->q21.1::q22.12--> q22.22::p13-->p13). The clinical signs can either be due to gene inactivation in connection with structural changes at the break and fusion regions, to the building of new fusion genes within the CCR and/or to the deletion of genes in 21q22.11. 相似文献
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Summary Three hundred and twenty-five aberrant breakpoints in chronic myelogenous leukaemia (CML) with Philadelphia chromosome variant were reviewed. Eight chromosomal bands (3p21, 6p21, 7p22, 11q13, 12p13, 17p13, 17q21, and 17q25) were found to be highly involved. Apart from 17q25, all these bands correspond to oncogenes sites and/or sites involved as primary breakpoints in cancer.This work was presented in part at the Congrès National d'Hématologie et de Transfusion Sanguine, March 1985 (Nouv Rev Fr Hématol, 1985, 27:72) and at the American Association for Cancer Research Meeting (Proceeding of the AACR 1986, 27:148) 相似文献
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Detection of amplified DNA sequences by reverse chromosome painting using genomic tumor DNA as probe
Stefan Joos Harry Scherthan Michael R. Speicher Jürgen Schlegel Thomas Cremer Peter Lichter 《Human genetics》1993,90(6):584-589
A modification of reverse chromosome painting was carried out using genomic DNA from tumor cells as a complex probe for chromosomal in situ suppression hybridization to normal metaphase chromsome spreads. Amplified DNA sequences contained in such probes showed specific signals, revealing the normal chromosome positions from which these sequences were derived. As a model system, genomic DNAs were analyzed from three tumor cell lines with amplification units including the proto-oncogene c-myc. The smallest amplification unit was about 90 kb and was present in 16–24 copies; the largest unit was bigger than 600 kb and was present in 16–32 copies. Specific signals that co-localized with a differently labeled c-myc probe on chromosome band 8q24 were obtained with genomic DNA from each cell line. In further experiments, genomic DNA derived from primary tumor material was used in the case of a male patient with glioblastoma multiforme (GBM). Southern blot analysis using an epidermal growth factor receptor gene (EGFR) probe that maps to 7p13 indicated the amplification of sequences from this gene. Using reverse chromosome painting, signals were found both on band 7p13 and bands 12q13–q15. Notably, the signal on 12q13–q15 was consistently stronger. The weaker 7p13 signal showed co-localization with the major signal of the differently labeled EGFR probe. A minor signal of this probe was seen on 12q13, suggesting cross-hybridization to ERB3 sequences homologous to EGFR. The results indicate co-amplification of sequences from bands 12q13–q15, in addition to sequences from band 7p13. Several oncogenes map to 12q13–q15 providing candidate genes for a tumor-associated proto-oncogene amplification. Although the nature of the amplified sequences needs to be clarified, this experiment demonstrates the potential of reverse chromosome painting with genomic tumor DNA for rapidly mapping the normal chromosomal localization of the DNA from which the amplified sequences were derived. In addition, a weaker staining of chromosomes 10 and X was consistently observed indicating that these chromosomes were present in only one copy in the GBM genome. This rapid approach can be used to analyze cases where no metaphase spreads from the tumor material are available. It does not require any preknowledge of amplified sequences and can be applied to screen large numbers of tumors. 相似文献
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Marie-Françoise Rousseau-Merck Josette Hillion Philippe Jonveaux Philippe Couillin Paule Seité Hans-Jürgen Thiesen Roland Berger 《Human genetics》1993,92(6):583-587
Nine KOX zinc finger genes were localized on four human chromosomes by in situ hybridization of cDNA probes to metaphase chromosomes. KOX1 (ZNF10), KOX11 (ZNF18), and KOX12 (ZNF19) were mapped to chromosome bands 12q24.33, 17p13-p12, and 16q22-q23, respectively. Six other KOX genes were localized on chromosome 19: KOX6 (ZNF14) and KOX13 (ZNF20) to 19p13.3-p13.2, KOX5 (ZNF13) and KOX22 (ZNF27) to 19q13.2-qter, and KOX24 (ZNF28) and KOX28 (ZNF30) to 19q13.4. Pulsed field gel electrophoresis experiments showed that the pairs of KOX genes found on the chromosome bands 12q24.33, 16q22-q23, 19p13.3-p13.2, or 19q13.3-qter lie within 200–300 kb DNA fragments. This suggests the existence of KOX gene clusters on these chromosomal bands. 相似文献
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Arginase deficiency manifesting delayed clinical sequelae and induction of a kidney arginase isozyme
Wayne W. Grody Rita M. Kern Deborah Klein Amy E. Dodson Paul B. Wissman Sanford H. Barsky Stephen D. Cederbaum 《Human genetics》1993,91(1):1-5
De novo chromosome structural abnormalities cannot always be diagnosed by the use of standard cytogenetic techniques. We applied a previously developed chromosome-band-specific painting method to the diagnosis of such rearrangements. The diagnostic procedures consisted of microdissection of an aberrant chromosomal region of a given patient, polymerase chain reaction (PCR) amplification of the dissected chromosomal DNA, and subsequent competitive fluorescence in situ hybridization (FISH) using the PCR products as a probe pool on metaphase chromosomes from the patient and/or a karyotypically normal person. With this strategy, we studied 6 de novo rearrangements (6p+, 6q+, 9p+, 17p+, +mar, and +mar) in 6 patients. These rearrangements had been seen by conventional banding but their origin could not be identified. In all 6 patients, we successfully ascertained the origin. Using an aberrant region-specific probe pool, FISH signals appeared on both the aberrant region and a region of another specific chromosome pair. A reverse probe pool that was generated through the microdissection of normal chromosomes at a candidate region for the origin of the aberration hybridized with both the aberrant and the candidate regions. We thus diagnosed one patient with 17p+ as having trisomy for 14q32-qter, one with 9p+ as having trisomy for 12pter-p12, one with 6q+ as having a tandem duplication (trisomy) of a 6q23-q25 segment, one with 6p+ as having a tandem duplication (trisomy) of a 6p23-q21.3 segment, one with a supernumerary metacentric marker chromosome as having tetrasomy for 18pter-cen, and the last with an additional small marker chromosome as having trisomy for 18p11.1 (or p11.2)-q11.2. The present targeted chromosome-band-painting method provides the simple and rapid preparation of a probe pool for region-specific FISH, and is useful for the diagnosis of chromosome abnormalities of unknown origin. 相似文献
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Molecular cytogenetic characterization of 17 rob(13q14q) Robertsonian translocations by FISH, narrowing the region containing the breakpoints. 总被引:3,自引:3,他引:0
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We have characterized 17 rob(13q14q) Robertsonian translocations, using six molecular probes that hybridize to the repetitive sequences of the centromeric and shortarm regions of the five acrocentric chromosomes by FISH. The rearrangements include six de novo rearrangements and the chromosomally normal parents, five maternally and three paternally inherited translocations, and three translocations of unknown origin. The D21Z1/D13Z1 and D14Z1/D22Z1 centromeric alpha-satellite DNA probes showed all rob(13q14q) chromosomes to be dicentric. The rDNA probes did not show hybridization on any of the 17 cases studied. The pTRS-47 satellite III DNA probe specific for chromosomes 14 and 22 was retained around the breakpoints in all cases. However, the pTRS-63 satellite III DNA probe specific for chromosome 14 did not show any signals on the translocation chromosomes examined. In 16 of 17 translocations studied, strong hybridization signals on the translocations were detected with the pTRI-6 satellite I DNA probe specific for chromosome 13. All parents of the six de novo rob(13q14q), including one whose pTRI-6 sequence was lost, showed strong positive hybridization signals on each pair of chromosomes 14 and 13, with pTRS-47, pTRS-63, and pTRI-6. Therefore, the translocation breakpoints in the majority of rob(13q14q) are between the pTRS-47 and pTRS-63 sequences in the p11 region of chromosome 14 and between the pTRI-6 and rDNA sequences within the p11 region of chromosome 13. 相似文献
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Deletion of chromosome 21 and normal intelligence: molecular definition of the lesion 总被引:7,自引:0,他引:7
Julie R. Korenberg Dagmar K. Kalousek Goran Anneren Stefan-M. Pulst Judith G. Hall Charles J. Epstein David R. Cox 《Human genetics》1991,87(2):112-118
Summary Application of a method for the fine structure analysis of unbalanced chromosomal rearrangements using quantitative Southern blot analysis has established that an individual of normal intelligence and largely normal appearance has a significant interstitial deletion of chromosome 21. Using high resolution cytogenetic analysis and molecular analysis with five single copy DNA sequences unique to chromosome 21 and a probe for human SOD1 (CuZn, Superoxide dismutase), we find that the deletion extends from the border of bands 21q11.1–11.2 and extends to the border of bands 21q21.2–q21.3. The latter border is established molecularly by the presence of two copies of SOD1, previously mapped to band 21q22.1, and of four single copy sequences known to be located distal to this region. The presence of SOD1 was confirmed by enzyme dosage analysis. These findings demonstrate that deletion of close to 20,000kb of autosomal material is compatible with normal intelligence. Further, they suggest that chromosome 21 may include a large region of relative developmental neutrality whose molecular basis may now be investigated. Because of the limits of even high resolution cytogenetic analysis, fine structure molecular analyses of this type will be necessary to reliably detect and define similar small chromosomal deletions or insertions. The molecular definition of such aneuploidy provides the basis for increasing the resolution of the human physical genetic map. 相似文献
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Nicholas Ho Hui Peng Chelsea Mayoh Pei Y. Liu Bernard Atmadibrata Glenn M. Marshall 《Cell cycle (Georgetown, Tex.)》2018,17(6):749-758
Neuroblastoma, the most common solid tumour in early childhood, is characterized by very frequent chromosomal copy number variations (CNVs). While chromosome 2p amplification, 17q gain, 1p and 11q deletion in human neuroblastoma tissues are well-known, the exact frequencies and boundaries of the chromosomal CNVs have not been delineated. We analysed the publicly available single nucleotide polymorphism (SNP) array data which were originally generated by the Therapeutically Applicable Research to Generate Effective Treatments (TARGET) initiative, defined the frequencies and boundaries of chromosomes 2p11.2 – 2p25.3 amplification, 17q11.1-17q25.3 gain, 1p13.3-1p36.33 deletion and 11q13.3-11q25 deletion in neuroblastoma tissues, and identified chromosome 7q14.1 (Chr7:38254795-38346971) and chromosome 14q11.2 (Chr14:21637401-22024617) deletion in blood and bone marrow samples from neuroblastoma patients, but not in tumour tissues. Kaplan Meier analysis showed that double deletion of Chr7q14.1 and Chr14q11.2 correlated with poor prognosis in MYCN gene amplified neuroblastoma patients. In conclusion, the oncogenes amplified or gained and tumour suppressor genes deleted within the boundaries of chromosomal CNVs in tumour tissues should be studied for their roles in tumourigenesis and as therapeutic targets. Focal deletions of Chr7q14.1 and Chr14q11.2 together in blood and bone marrow samples from neuroblastoma patients can be used as a marker for poorer prognosis and more aggressive therapies. 相似文献
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Thirteenfold increase of chromosomal aberrations non-randomly distributed in chagasic children treated with nifurtimox 总被引:3,自引:0,他引:3
Chromosomal aberrations were analyzed from cultures of peripheral lymphocytes in 2 groups of chagasic children, before and after treatment with nifurtimox. The mean incidence of chromosomal aberrations increased from control values of 1.75 +/- 1.39 (8 patients) to 23.55 +/- 9.55 (6 patients) at a significance of P less than 0.0001. G-banding analysis of chromosomal aberration sites revealed that treated patients present coincidence in the chromosome regions affected: 1p11, 1q11-12, 9q11-13, 17q11-21, 2p21, 2q23, 2q31, 2q33, 6p21, 6p21, 7q32, 13q14, 13q22, 15q22. These data indicate a non-random distribution of chromosomal aberrations induced by nifurtimox therapeutic treatment. 相似文献
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This review summarizes the chromosomal changes detected by molecular cytogenetic approaches in esophageal squamous cell carcinoma (ESCC), the ninth most common malignancy in the world. Whole genome analyses of ESCC cell lines and tumors indicated that the most frequent genomic gains occurred at 1, 2q, 3q, 5p, 6p, 7, 8q, 9q, 11q, 12p, 14q, 15q, 16, 17, 18p, 19q, 20q, 22q and X, with focal amplifications at 1q32, 2p16-22, 3q25-28, 5p13-15.3, 7p12-22, 7q21-22, 8q23-24.2, 9q34, 10q21, 11p11.2, 11q13, 13q32, 14q13-14, 14q21, 14q31-32, 15q22-26, 17p11.2, 18p11.2-11.3 and 20p11.2. Recurrent losses involved 3p, 4, 5q, 6q, 7q, 8p, 9, 10p, 12p, 13, 14p, 15p, 18, 19p, 20, 22, Xp and Y. Gains at 5p and 7q, and deletions at 4p, 9p, and 11q were significant prognostic factors for patients with ESCC. Gains at 6p and 20p, and losses at 10p and 10q were the most significant imbalances, both in primary carcinoma and in metastases, which suggested that these regions may harbor oncogenes and tumor suppressor genes. Gains at 12p and losses at 3p may be associated with poor relapse-free survival. The clinical applicability of these changes as markers for the diagnosis and prognosis of ESCC, or as molecular targets for personalized therapy should be evaluated. 相似文献