水仙荧光原位杂交体系的建立     

吴菁华  张志忠  吕柳新 《勤云标准版测试》2008,(11)

建立以水仙染色体为靶、rDNA为探针的荧光原位杂交实验技术,为进一步利用荧光原位杂交技术分析水仙的亲缘和进化关系奠定了基础.水仙的染色侉制片以去壁低渗-火焰干燥法较好,容易获得大量清晰、分散的有丝分裂中期相;切口平移法地高辛标记探针、染色体和探针90℃变性5min能有效的进行水仙rDNA的染色体荧光原位杂交定位.  相似文献   

棉花gDNA体细胞染色体FISH技术   总被引：17，自引：6，他引：11  

王春英  王坤波  王文奎  李懋学  宋国立  崔荣霞  黎绍惠  张香娣  张家明 《棉花学报》1999,(2)

介绍了棉花基因组ＤＮＡ（ｇｅｎｏｍｅＤＮＡ,简为ｇＤＮＡ）体细胞染色体荧光原位杂交〔ＦＩＳＨ〕的技术流程,并着重分析和讨论了影响试验结果的关键因素,包括染色体和探针的变性条件、染色体的蛋白酶Ｋ处理技巧等。试验中作为靶ＤＮＡ的体细胞染色体采用棉属异源四倍体种海岛棉;探针和封阻均采用ｇＤＮＡ,材料是棉属二倍体种Ａ染色体组（Ａｇｅｎｏｍｅ）的棉种（亚洲棉和草棉）和Ｄ染色体组（Ｄｇｅｎｏｍｅ）的棉种（瑟伯氏棉、雷蒙德氏棉、戴维逊氏棉等）,分别交互使用。试验结果比较理想,获得良好的ＦＩＳＨ片子,而且重复性好。  相似文献   

半滑舌鳎含sox9基因BAC克隆的筛选及BAC-FISH定位          下载免费PDF全文

孙建  陈松林  谢明树  邵长伟 《渔业科学进展》2014,35(1):34-40

以本实验室构建的半滑舌鳎BAC文库为基础,通过对其进行有序混合,构建了两步PCR筛选体系;对性别决定相关的sox9基因的序列设计特异性引物,筛选获得阳性单一BAC克隆。利用BAC-FISH技术将包含sox9基因的BAC克隆定位在半滑舌鳎染色体上。结果显示,sox9基因在雌、雄半滑舌鳎的一对常染色体的长臂上分别存在两个杂交信号位点,信号稳定且特异。研究证实了该文库筛选体系的有效性;首次实现了对性别相关的sox9基因在半滑舌鳎染色体上的定位,其结果为揭示sox9基因参与鱼类性别控制的机制提供了重要基础。  相似文献   

Screening and Positioning of Three Chromosome-specific BAC Clones in Gossypium raimondii   总被引：1，自引：1，他引：0  

QIN 　Qin  LIU 　Fang  GAN Yi-Mei  WANG Chun-Ying  WANG Yu-Hong  CAI Xiao-Yan  WANG Kun-Bo 《棉花学报》2013,25(4):323-328

We screened a bacterial artificial chromosome(BAC) library of Gossypium barbadense acc. Pima 90-53 to identify chromosome-specific BAC clones. Using BAC-fluorescence in situ hybridization technology, we obtained three BAC clones specific to chromosome D₅01, D₅02, and D₅10 of Gossypium raimondii, which could be used as cytological markers for those three chromosomes. Comparative mapping of these three BACs between G. barbadense and in G. raimondii showed that these three BAC clones could also be used to identify chromosomes D_b01, D_b02, and D_b10 in G. barbadense. The position of the BAC clone 280G06 on chromosome 10 did not show colinearity between G. barbadense and G. raimondii, possibly because of chromosomal rearrangement.  相似文献   

The pig CART (cocaine- and amphetamine-regulated transcript) gene and association of its microsatellite polymorphism with production traits     

M. Stachowiak  J. Cieslak  A. Skorczyk  J. Nowakowska  I. Szczerbal  M. Szydlowski  & M. Switonski 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2009,126(1):37-42

  相似文献   

An oligonucleotide probe for the detection of Erwinia herbicola and Erwinia ananas     

Yoshihiro Nakanishi  Appolinaire Adandonon  Ikuko Okabe  Yuko T. Hoshino  Naoyuki Matsumoto 《Journal of General Plant Pathology》2006,72(5):328-333

An oligonucleotide probe targeting the rRNA of Erwinia herbicola and Erwinia ananas was designed to detect their cells using fluorescence in situ hybridization. The Cy3-labeled probe hybridized strongly with these species but very weakly with nontarget species such as Erwinia mallotivora, Erwinia nigrifluens, Erwinia cypripedii, Erwinia chrysanthemi, Erwinia carotovora subsp. carotovora, E. carotovora subsp. atroseptica, and Escherichia coli. This technique visualized E. herbicola cells after inoculation of kumquat fruits. The probe is promising as a tool for studying population dynamics of E. herbicola and E. ananas.  相似文献   

洋葱染色体核型的FISH分析   总被引：2，自引：0，他引：2  

赵泓  O.Schrader  R.Ahne 《园艺学报》2003,30(4):463-464

 挑选洋葱(Allium cepa L．) (2n=2x=16，CC)有丝分裂中期细胞，以洋葱卫星DNA序列(AC—SAT-DNA)为探针，DIG-1 1-dUTP为标记物，用荧光原位杂交(FISH)方法观察其在染色体上的定位，并进行洋葱的核型分析。6号染色体为近端部着丝点染色体，在其长臂末端有一个卫星DNA序列的主要位点。其余每条染色体在其长、短臂的末端都各具有一个主要位点。差异表现在各染色体上卫星DNA序列的次要位点分布不同。根据洋葱8对染色体各自具有的独特的卫星DNA分布位点，可将洋葱8对染色体全部分开。以此为基础，构建了洋葱卫星DNA在染色体上分布的模式图。  相似文献   

Gastric Helicobacter species as a cause of feline gastric lymphoma: a viable hypothesis     

Bridgeford EC  Marini RP  Feng Y  Parry NM  Rickman B  Fox JG 《Veterinary immunology and immunopathology》2008,123(1-2):106-113

Gastric Helicobacter spp. are associated with chronic inflammation and neoplastic transformation in humans as well as domestic and laboratory species. The present study examined the association of Helicobacter heilmannii (Hhe) infection in pet cats with feline gastric mucosa associated lymphoid tissue (MALT) lymphoma. Tissues were collected via gastric biopsy or at necropsy from 47 pet cats with clinical signs of gastrointestinal disease, including vomiting and inappetance, and classified as gastritis (14/47), lymphoma (31/37), or normal (2/47). Tissues positive for argyrophilic organisms with Warthin–Starry stain (29/47) were assessed by fluorescent in situ hybridization (FISH) for the presence of Hhe strains 1–4 as well as with a fifth probe that detected Helicobacter salomonis, Helicobacter bizzozeronii, or Helicobacter felis. A significant association of positive Warthin–Starry status with Hhe infection was found in cases of sick cats (22/29; p < 0.05 by Chi-square; χ² = 7.034). Interestingly, a significant association between Hhe status and a diagnosis of lymphoblastic or lymphocytic lymphoma was observed as well in a subset of 24 Warthin–Starry positive lymphoma cases: of lymphoblastic lymphoma cases, 13/17 were positive for Hhe (p < 0.05; χ² = 4.854). Hhe strains 2 and 4 were most commonly found (18/29 and 17/29, respectively) among sick cats, although a higher than expected number of cats was also positive for Hhe1, which initial reports have described as rare in cats and common in humans. The association found between a positive Hhe status with the presence of feline gastric lymphoma, especially lymphoblastic lymphoma, argues for the need to conduct prospective studies to better identify the frequency and strain distribution of Hhe infection in both healthy and clinically ill cats, particularly those cats with gastric lymphoma.  相似文献   

家蚕HSP20·8基因体外表达及荧光原位杂交研究(英文)     

孙卫忠  李斌  王彦文  柴春利  刘彬斌  鲁成 《蚕业科学》2007,33(1):14-19

前期研究发现,相对于其它组织器官,有一种低分子量热激蛋白在家蚕肾型卵中高量表达,并命名为低分子量热激蛋白HSP20.8。为了进一步阐明家蚕低分子量热激蛋白HSP20.8的基因结构、表达特性及其在家蚕染色体上的分布,根据HSP20.8基因cDNA序列设计特异引物,进行了克隆、表达及荧光原位杂交研究。结果表明,该基因含有1个561碱基对的开放阅读框(open reading frame,ORF),与cDNA序列比较分析发现该基因无内含子序列。重组蛋白的分子量在20 kD左右。热激蛋白基因HSP20.8在家蚕基因组中为单拷贝基因,其位点在染色体近中部区域。  相似文献   

FISH mapping of 10 canine BAC clones harbouring genes and microsatellites in the arctic fox and the Chinese raccoon dog genomes     

I. Szczerbal  J. Klukowska-Roetzler  G. Dolf  C. Schelling  & M. Switonski 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2006,123(5):337-342

Cytogenetic mapping of the arctic fox and the Chinese raccoon dog were performed using a set of canine probes derived from the Bacterial Artificial Chromosome (BAC) library. Altogether, 10 BAC clones containing sequences of selected genes (PAX3, HBB, ATP2A2, TECTA, PIT1, ABCA4, ESR2, TPH1, HTR2A, MAOA) and microsatellites were mapped by fluorescence in situ hybridization (FISH) experiments to chromosomes of the canids studied. At present, the cytogenetic map on the arctic fox and Chinese raccoon dog consists of 45 loci each. Chromosomal localization of the BAC clones was in agreement with data obtained by earlier independent comparative chromosome painting. However, two events of telomere‐to‐centromere inversions were tentatively identified while compared with assignments in the dog karyotype.  相似文献