Development of new spectral karyotyping (SKY) techniques in cytogenetic analysis

细胞遗传学分析中新型光谱核型分析 (SKY) 技术的开发

• 批准号: 11671010
• 负责人: KAKAZU Naoki
• 金额: $ 1.66万
• 依托单位: Kyoto Prefectural University of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671010/
• 关键词: cytogenetic analysis; G-banding; SKY; dual karyotyping; color-banding; color-banding; multicolor-banding法; microdissection

1. dual karyotypingThe DAPI-band image, which was digitally inverted and contrast-enhanced, was almost identical to the pattern obtained with G-banding. This method resulted in adequate band resolution that corresponded to approximately 400 bands per haploid set. The karyotypes (dual karyotypes) were ascertained by combining SKY and the DAPI-banding, in which both the DAPI-band image and the classification color image of the chromosomes are simultaneously displayed in parallel. The karyotype helped determine the origin of the rearranged chromosomal materials by using the spectrally classified color images. In addition, the breakpoints could be precisely determined with respect to chromosome bands on the G-band-like images.2. color-bandingWe developed a novel chromosome banding technique named spectral color banding (SCAN). With this technique we displayed a multicolor banding pattern that almost entirely correlated with the corresponding G-banding pattern. SCAN analysis allowed for accurate identification of chromosome-band origin on the basis of its spectral color and detection of intrachromosomal changes. Furthermore, SCAN analysis could determine the precise breakpoints of complex rearrangements and identify the chromosome-band origin of small chromosome segments. Our preliminary use of SCAN in combination with SKY suggests that this new technique is capable of accurately identifying chromosome-band origin for each of the 24 human chromosomes, and that it may be able to identify intrachromosomal changes not previously detected by G-banding or SKY.SCAN technique could become a powerful tool for cytogenetic analysis and an important step on the road to fully automated karyotyping.

1. DAPI-band图像，经过数字反转和对比度增强，与g -band图像几乎相同。这种方法产生了足够的波段分辨率，对应于每个单倍体集大约400个波段。将SKY和dapi带带结合，同时平行显示dapi带带图像和染色体的分类彩色图像，确定了染色体的核型（双核型）。核型通过光谱分类彩色图像帮助确定重排染色体物质的起源。此外，在g带样图像上，可以精确地确定染色体带的断点。我们开发了一种新的染色体条带技术，称为光谱彩色条带（SCAN）。通过这种技术，我们显示了一个几乎完全与相应的g带模式相关的多色带模式。扫描分析允许在其光谱颜色和检测染色体内的变化的基础上，染色体带起源的准确鉴定。此外，SCAN分析可以确定复杂重排的精确断点，并确定小染色体片段的染色体带起源。我们将SCAN与SKY结合的初步使用表明，这项新技术能够准确地识别24条人类染色体中的每一条染色体的染色体带起源，并且它可能能够识别以前未被g带或SKY检测到的染色体内变化。扫描技术将成为细胞遗传学分析的有力工具，是迈向全自动核型的重要一步。

项目成果

Kakazu N, et al.: "Characterization of complex chromosomal abnormalities in B-cell lymphoma by a combined spectral karyotyping (SKY) analysis and fluorescence in situ hybridization (FISH) using a 14q telomere probe"Am J Hematol. 65. 291-297 (2000)

Kakazu N 等人：“使用 14q 端粒探针，通过组合光谱核型分析 (SKY) 和荧光原位杂交 (FISH) 来表征 B 细胞淋巴瘤中复杂的染色体异常”Am J Hematol。

阿部達生: "造血器腫瘍アトラス"日本医事新報社. 284 (2000)

阿部达雄：《造血肿瘤图谱》《日本医治新报社》284（2000）。

Chu D, et al.: "Cloning and characterization of LUN, a novel RING-finger protein that is highly expressed in lung and specifically binds to a palindromic sequence."J Biol Chem. 276. 14004-14013 (2001)

Chu D 等人：“LUN 的克隆和表征，LUN 是一种在肺中高度表达并特异性结合回文序列的新型环指蛋白。”J Biol Chem。

Minakuchi M, et al.: "Identification and characterization of SEB, anovel protein that binds to the acute undifferentiated leukemia-associated protein SET"Eur J Biochem. 268. 1340-1351 (2001)

Minakuchi M 等人：“SEB 的鉴定和表征，SEB 是一种与急性未分化白血病相关蛋白 SET 结合的新型蛋白”，Eur J Biochem。

Nosaka K, et al.: "Isolation and characterization of a human thiamine pyrophosphokinase cDNA."Biochim Biophys Acta. 1517. 293-297 (2001)

Nosaka K 等人：“人硫胺素焦磷酸激酶 cDNA 的分离和表征。”Biochim Biophys Acta。