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1.
采用原位杂交和RNA点杂交方法,观察了一种新的端粒相关锌指基因TASL30在早期人胚与50种器官组织中的mRNA表达。结果表明,TASL30基因在早期人胚神经管中有明显表达,其中在神经管的头端表达最强,而在人的50种器官组织中均未检测到该基因的明显表达。另外通过G-显带和染色体原位杂交,将TASL30基因定位于人染色体12q24~qter。 Abstract:By means of in situ hybridization and RNA dot blot,mRNA expression of a telomereic-associated zinc finger gene(TASL30 )was observed.The results showed that TASL30 gene was expressed in significant amount in neural tube of early human embryo and the cephalic expression level was the highest,but none of recognizable positive signal was detected in 50 human tissues.By G-banding and chromosome in situ hybridization,TASL30 gene was located in chromosome 12q24~qter.  相似文献   

2.
冯耀东  高立  苟德明  蒋达和  毛歆  李文鑫 《遗传》2000,22(6):361-364
采用原位杂交和RNA点杂交方法,观察了一种新的端粒相关锌指基因TASL30在早期人胚与50种器官组织中的mRNA表达。结果表明,TASL30基因在早期人胚神经管中有明显表达,其中在神经管的头端表达最强,而在人的50种器官组织中均未检测到该基因的明显表达。另外通过G-显带和染色体原位杂交,将TASL30基因定位于人染色体12q24~qter。 Abstract:By means of in situ hybridization and RNA dot blot,mRNA expression of a telomereic-associated zinc finger gene(TASL30 )was observed.The results showed that TASL30 gene was expressed in significant amount in neural tube of early human embryo and the cephalic expression level was the highest,but none of recognizable positive signal was detected in 50 human tissues.By G-banding and chromosome in situ hybridization,TASL30 gene was located in chromosome 12q24~qter.  相似文献   

3.
应用荧光原位杂交(FISH)技术研究 黑叶猴染色体易位   总被引:5,自引:0,他引:5  
本文应用染色体荧光原位杂交(FISH)技术,利用人9号和14号染色体特异探针,对深低温冻存和长期传代的黑叶猴细胞株染色体畸变进行了分析。确定在长期冻存和传代过程中,一些黑叶猴细胞在No.12和No.17染色体之间发生了易位,一条 No.17染色体发生断裂,断裂点在17q13,断裂片段17q13-17qter易位到一条 No.12染色体长臂末端,形成一条小的中着丝粒的和一条具较长长臂的衍生染色体即 der(17) 和 der(12)。结果表明,荧光原位杂交技术用人染色体特异探针不仅能检测出人类染色体畸变,也能有效地检测灵长类动物染色体畸变。 Abstract In this paper,the chromosome aberration of long-term cryopreserved and subcultured Francois' monkey (Semnopithecus francoisi) cell line(KCB 92008) was analyzed by fluoresence in situ hybridizaton (FISH) using human 9 and 14 chromosome DNA probes. After compared the hybridization pattern with the G-banding pattern on the same metaphase,a translocation between Nos.12 and 17 chromosomes was identified. In some Francois'monkey cells,one of chromosome No.17 was broken into two at the breakpoint 17q13,the segment(17q13-17qter) without centromere transfered to the long arm terminal of one chromosome No.12. Thus,two derivant chromosomes der(12) and der(17) were formed,the long arm of der(12) was longer than the normal partner,while the long arm of der(17) was shorter than the normal one. The result indicated that the technique of FISH using human whole chromosome probes was not only a powerful tool to detect human chromosome rearrangements,but also a usefulmethod to study the primate chromosome aberration.  相似文献   

4.
玉米cyclinⅢ基因的染色体原位杂交物理定位   总被引:2,自引:0,他引:2  
本文首次报道了玉米低拷贝基因 cyclinⅢ(B-类)生物素标记的染色体原位杂交定位结果。供试探针为该基因的cDNA克隆,其长度仅为1.6kb。结果表明, 探针的信号分布在第6染色体短臂和第9染色体长臂,与着丝粒的百分距离分别为70.05±3.31和86.86±1.64,检出率分别为8.29%和6.83%。文中对基因的物理位置与功能间的关系等作了讨论。 Abstract:A biotin-labelled in situ hybridization technique was used to physically map a low copy gene cyclinIII in maize.The cDNA clone was 1.7kb in size.The probe was hybridized onto the short arm of chromosome 6 and the long arm of chromosome 9.The percent distances from centromere to detection site were 70.05±3.31 and 86.86±1.64 respetively.The detection rates of in situ hybridization were 8.29 and 6.83 respectively,The relationship between the position and function of the genes is discussed in this paper.  相似文献   

5.
Study on gene sensor based on primer extension   总被引:1,自引:0,他引:1  
Based on the fact that the resonant frequency of a piezoelectric crystal is the function of its surface deposit, and that the primer extends after it hybridizes with the template, the primer extension gene sensor technique was developed. The prominent feature of the technique is that fast and sensitive frequency signals are used as the monitoring system of gene hybridization and primer strand extension. Results show that this technique may be used in homologous analysis of nucleic acid, trace DNA detection, and determining the integration of DNA. It may also be used for isolation of target gene, gene mutation analysis, and predicting the location of a gene in its genome.  相似文献   

6.
用FISH技术对人、恒河猴、食蟹猴染色体的研究   总被引:6,自引:1,他引:6  
用人类5号、 9号、13号、15号、17号、20号整条染色体探针分别对人、恒河猴和食蟹猴的中期细胞进行荧光原位杂交,结果表明:人的5号、13号、17号探针分别杂交到恒河猴的5号、16号、17号染色体上;9号探针杂交到恒河猴14号染色体的长臂及部分短臂上; 15号探针杂交到恒河猴7号染色体短臂及部分长臂上;20号探针杂交到恒河猴的13号染色体长臂上。食蟹猴的杂交结果与恒河猴完全一致。结合G带带型分析,对人与猕猴的染色体同源性及其进化进行了讨论。 Abstract:Fluorescent in situ hybridizaiton(FISH)was used on the metaphase of Macaca mulatta and Macaca fasicularis with human chromosome specific DNA libraries for chromosome 5、9、13、15、17 and 20.In Macaca mulatta,the result showed that chromosome 5、16 and 17 was entirely painted by human chromosome 5、13 and 17 specific libraries respectively.The long arm and the partial short arm of chromosome 14 and the short arm and the partial long arm of chromosome 7 were painted by human chromosome 9 and 15 specific libraries respectively.And the long arm of chromosome 13 was painted by human chromosome 20 library.The result was the same in Macaca fasicularis.Combinded with the comparative analysis of G-banding,the evolutional relationship of these chromosomes between human and macaques was discussed.  相似文献   

7.
为了确定两例细胞遗传学提示染色体结构异常的核型,应用通过显微切割技 术构建的人类18号和7号染色体探针池,分别对这两例病例的中期分裂相进行染色体涂染,结合显带染色体,确定两者核型分别为46,XY,t(3;18) (q12;q21)和46,XX,dir ins(1;7)(p3104;q34q36)。染色体涂染技术是染色体显带技术的重要补充和发展,为染色体结构异常提供了一种直观、准确的检测手段,在遗传咨询和产前诊断方面有重要作用。 Abstract:In this study,chromosome painting technique was performed to analyse the abnormal karyotypes of two carriers.Chromosome 18 and 7 specific libraries,which were generated by chromosome microdissection technique,were used as probe pools to hybridize the carriers metaphase chromosomes respectively.Unlabled human genomic DNA was used to inhibit the hybridization of sequences in the library that bind to mutiple chromosomes.Structure abnormality was detected clearly in metaphase.Combined with the banding chromosomes,we concluded that their karytypes were 46,XY,t(3;18)(q12;q21)and 46,XX,dir ins(1;7)(p3104;q34q36).Chromosome painting,as a direct and concise method in analysing chromosome structure abnormality,is an important complement and development of chromosome banding technique,and has important application in genetic counselling and prenatal diagnosis.  相似文献   

8.
Chen K  Li D  Jiang YH  Yao WJ  Wang XJ  Wei XC  Gao J  Xie LD  Yan ZY  Wen ZY  Chien S 《Cell research》2004,14(2):161-168
The cDNA fragment of human TRAIL (TNF-related apoptosis inducing ligand) was cloned into RevTet-On, a Tetregulated and high-level gene expression system. The gene expression system was constructed in a human leukemic cell line: Jurkat. By using RevTet-On TRAIL gene expression system in Jurkat as a cell model, we studied the influence of TRAIL gene on the changes of cellular apoptosis before and after the TRAIL gene expression, which was induced by adding tetracycline derivative doxycycline (Dox). The results indicated that the cellular apoptosis ratio was largely dependent on the TRAIL gene expression level. Moreover, it was found that the apoptosis-inducing TRAIL could cause significant changes in the biophysical properties of Jurkat cells. The cell surface charge density decreased, the membrane fluidity declined, the elastic coefficients K1 increased, and the proportion of α-helix in membrane protein secondary structure decreased. Thus, the apoptosis-inducing TRAIL gene caused significant changes on the biomechanic properties of Jurkat cells.  相似文献   

9.
An efficient plasmid transformation system for S. mycarofaciens 1748 has been established. In order to determine the function of MKR gene in S. mycarofaciens 1748, the gene disruption experiment was carried out. For this purpose the plasmid pKC1139 was used. A recombinant strain with white spore appeared, in contrast to the grey-colour spore of S. mycarofaciens 1748. This suggested that homologous recombination between plasmid-borne MKR gene sequence and the chromosome of S. mycarofaciens 1748 had occurred. A Southern hybridization experiment using α- P-labelled MKR gene as probe indicated that the desired integration event had occurred in the re-combinant. The result of gene disruption showed that the alteration of this gene in the chromosome of S. mycarofa-ciens 1748 made sporulating colonies remain white instead of taking on the typical grey colour of sporulating wild type colonies, suggesting that MKR gene is involved in the biosynthesis of a spore pigment. The recombinant strain was in-cubated wit  相似文献   

10.
11.
In humans, the poly(A)-binding proteins (PABPs) comprise a small nuclear isoform and a conserved gene family that displays at least three functional proteins: PABP1, inducible PABP (iPABP), and PABP3, plus four pseudogenes (1, 2, 3, and PABP4). In situ hybridization of PABP3 cDNA as the probe on metaphasic chromosomes have revealed five possible loci for this gene family at 2q21-q22, 13q11-q12, 12q13.3-q15, 8q22, and 3q24-q25. Amplifications of specific DNA fragments from a human-rodent somatic cell hybrid panel have allowed us to associate PABP1 and PABP3 with 8q22 and 13q11-q12, respectively. The iPABP gene has been assigned to chromosome 1. This result, compared with radiation hybrid database information, strengthens the location of this gene to 1p32-p36. The pseudogenes PABP4, 1, and 2 have been assigned to chromosomes 15, 4, and 14, respectively. Three loci detected on chromosome spreads are not associated with any amplified fragment. They might represent other related PABP genes not yet identified.  相似文献   

12.
Chromosome 13q has been suggested as the site of a gene predisposing to human breast cancer, because loss of heterozygosity of alleles on this chromosome has been observed in some ductal breast tumors and because two breast cancer lines are altered at the retinoblastoma gene (RB1) at 13q14. To test this possibility, linkage of breast cancer susceptibility to 14 loci on chromosome 13q loci was assessed in extended families in which breast cancer is apparently inherited as an autosomal dominant trait. RB1 was excluded as the site of a breast cancer gene by a lod score of Z = -7.60 at close linkage for 13 families. Multipoint analysis yielded negative lod scores throughout the region between 13q12 and 13q34; over most of this distance, Z less than -2.0. Therefore, chromosome 13q appears to be excluded as the site of primary lesion for breast cancer in these families. In addition, comparison of tumor versus normal tissues of nonfamilial breast cancer patients revealed an alteration at the 5' end of RB1 in a mucoid carcinoma but no alterations of RB1 in five informative ductal adenocarcinomas. Linkage data and comparisons of tumor and normal tissues suggest that changes in the RBI locus either are secondary alterations associated with progression of some tumors or occur by chance.  相似文献   

13.
Atopic dermatitis (AD) is a common, itchy skin disease of complex inheritance characterized by dermal and epidermal inflammation. The heritability is considerable and well documented. To date, four genome scans have examined the AD phenotype, showing replicated linkage at 3p26-22, 3q13-21 and 18q11-21. Our previous AD scan showed evidence of linkage to loci at 3p and 18q, and furthermore at 4p15-14. In order to further investigate the genetic basis of AD, we collected and analysed a new Danish family sample consisting of 130 AD sib pair families (555 individuals including 295 children with AD). AD was diagnosed after clinical examination, AD severity was scored and specific IgE was determined. A linkage scan of chromosome 3, 4 and 18 was performed using 91 microsatellite markers. Linkage analyses were performed of dichotomous phenotypes and semi-quantitative traits including the AD severity score. We analysed the novel AD sample alone and together with the previously examined sample. AD severity showed a maximum Z-score of 3.7 at 4q22.1 suggesting the localization of a novel gene for AD severity. A maximum MOD score of 4.6 was obtained at 3p24 for the AD phenotype, providing the first significant linkage of AD at this locus. A maximum MLS score of 3.3 was obtained at 3q21 for IgE-associated AD, and evidence of linkage was also obtained at 3p22.2-21.31, 3q13, 4q35, and 18q12. The results presented should provide a firm basis for gene-targeting studies of AD and related disorders.  相似文献   

14.
15.
A novel Drosophila melanogaster gene UBL3 was characterized and shown to be highly conserved in man and Caenorhabditis elegans (C. elegans). The human and mouse homologues were cloned and sequenced. UBL3 is a ubiquitin-like protein of unknown function with no conserved homologues in yeast. Mapping of the human and mouse UBL3 genes places them within a region of shared gene order between human and mouse chromosomes on human chromosome 13q12-13 and telomeric mouse chromosome 5 (MMU5).  相似文献   

16.
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   

17.
Summary We describe the cytogenetic findings in a recurrent neurofibrosarcoma in a patient with nonfamilial von Recklinghausen disease. The composite karyotype was: 40,Y,-X,+dic r(X;20)(:Xp22.2q26::20p13 q13:), -1, +der(1)t(1;3) (p21;p24),-3,-4,-5,+der(5) t(5;?)(q31;?),-9,-9,+der(9)t(3;9)(q21 or q13;p24 or p22), -11,+der(11)t(11;?)(q22.2;?), -17,+der(17)t(17; 22;?)(q21;q13.1;?), -20, -21, -22, -22, +der(22)t(17; 22;?)(q21;q13.1;?),t(2;10)(q37;q22). The derivative chromosomes were demonstrated at the 500 band level. Chromosomes 17 and 22 were shown to be involved in an unbalanced three-way translocation: t(17;22;?)(q21;q13.1;?). This event was confirmed by in situ hybridization, using two probes mapped to chromosome 17. Hill H is a probe derived from the novel oncogene TRE and is located at 17q12–22. The second probe, derived from the granulocyte colony-stimulating factor (G-CSF), is located at 17q11–q21. The rearrangement between chromosomes 17 and 22 showed breakpoints similar or close to the gene loci for neurofibromatosis 1 (NF-1) and NF-2. Based on our observations we recommend that genetic studies on NF-1 tumors include both gene sites (NF-1 and NF-2) rather than focus on one gene locus.  相似文献   

18.
Maternal duplications of the imprinted 15q11-13 domain result in an estimated 1%-2% of autism-spectrum disorders, and linkage to autism has been identified within 15q12-13. UBE3A, the Angelman syndrome gene, has, to date, been the only maternally expressed, imprinted gene identified within this region, but mutations have not been found in autistic patients. Here we describe the characterization of ATP10C, a new human imprinted gene, which encodes a putative protein homologous to the mouse aminophospholipid-transporting ATPase Atp10c. ATP10C maps within 200 kb distal to UBE3A and, like UBE3A, also demonstrates imprinted, preferential maternal expression in human brain. The location and imprinted expression of ATP10C thus make it a candidate for chromosome 15-associated autism and suggest that it may contribute to the Angelman syndrome phenotype.  相似文献   

19.
The human MEP1A gene encodes the meprin alpha subunit that consists of a protease domain conserved in the astacin family of metalloendopeptidases and several C-terminal interaction domains present in other proteins. Using the alpha subunit cDNA, we identified two clones from a human P1-derived artificial chromosome (PAC) library. Fluorescence in situ hybridization (FISH) mapped both PACs (1e12, 65a14) to chromosome 6p21, confirming the MEP1A location. FISH also mapped PAC 65a14 to chromosome 13cen, and to chromosome 9 in three different regions, 9p12-13, 9q21, and 9q22. Southern blot analysis showed that sequences of PAC 65a14 and MEP1A were similar in the 3' end but different in the 5' end, revealing for the first time that the human genome may encode multiple interaction domains highly similar to those of the meprin alpha subunit. The symbols of MEP1AL1, MEP1AL2, MEP1AL3, and MEP1AL4 have been designated for MEP1A-like sequences on 9p12-13, 9q21, 9q22, and 13cen, respectively.  相似文献   

20.
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