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Featured researches published by Tetsuya Hirota.


Human Genetics | 1992

Chromosome-band-specific painting : chromosome in situ suppression hybridization using PCR products from a microdissected chromosome band as a probe pool

Han Xiang Deng; Koh-ichiro Yoshiura; Roeland W. Dirks; Naoki Harada; Tetsuya Hirota; Kazuhiro Tsukamoto; Yoshihiro Jinno; Norio Niikawa

SummaryWe describe a chromosome-band-specific painting method that involves (1) microdissection of the chromosome, chromosomal region or band, (2) amplification of a variety of chromosome/region/band-specific DNA fragments with the polymerase chain reaction (PCR), and (3) chromosome in situ suppression hybridization (CISS) with the direct use of the PCR products as a probe pool. With this method, it was possible 1) to paint an entire X or Y chromosome, a distal one-fourth of 2q, and only a band at 8q24.1, 2) to identify the origin of a minute marker chromosome in a mentally retarded patient, 3) to detect an X;Y translocation in another patient, and 4) to identify one human chromosome 2 in a human-mouse hybrid cell line. This method allows us to identify not only structural chromosome abnormalities at the band level, but also the origin of cytogenetically unidentifiable marker chromosomes. It will also be useful in studies of evolutionary cytogenetics.


Journal of Human Genetics | 1989

Satellited chromosome 9 in a boy with multiple anomalies

Naoki Harada; Kyohko Abe; Tatsuro Kondoh; Tetsuya Hirota; Norio Niikawa

SummaryA 5-year-old boy with multiple congenital anomalies showed a satellited long-arm chromosome 9, a previously undescribed abnormality. Various banding analyses of his chromosomes and those of his parents indicated that a reciprocal translocation, t(9;22)(q34.3;q11.21), occurred in the fathers gonad, and one of the translocation chromosomes was then transmitted to the patient. Thus, the patients karyotype was interpreted as 46,XY,-9,+psudic(9),t(9;22)(q34.3;q11.21). He showed several features similar to those of the Williams syndrome. The gene(s) responsible for the syndrome thus could be at either 9q34.3-qter or 22pter-q11.2. Southern blot analysis of the patients DNA indicated the presence of two copies of the argininosuccinate synthetase gene which had been assigned to 9q34.1-qter. In view of the fact that the 9q34.3-qter segment is monosomic in the patient, the gene locus was deduced to be at 9q34.1-q34.2 segment.


Journal of Human Genetics | 1992

Isolation of 2 novel RFLP markers and their localization at 2q35 by microdissection and subsequent enzymatic amplification

Tetsuya Kibe; Kazuhiro Tsukamoto; Tetsuya Hirota; Koh-ichiro Yoshiura; Tohru Ohta; Takaya Tohma; Toshiya Tamura; Yoshihiro Jinno; Norio Niikawa

SummaryWe previously constructed a chromosome 2q-specific genomic library and isolated a number of microclones. In the present study, we first analyzed with Southern hybridization whether any of the microclones represent restriction fragment length polymorphisms (RFLPs) and then tried to map RFLP markers physically, using the recently developed chromosome microdissection/enzymatic amplification method. Of 13 clones analyzed, two were RFLP markers; a clone, pM2C83, showed a fourallele MspI RFLP, and the other, pM2C8, a two-allele RsaI RFLP. In order to assign the two polymorphic markers, two chromosomal segments, 2q32-q35 and 2q35-qter, on the chromosome 2 from a karyotypically normal person were microdissected, and the DNA from each segment was amplified with the polymerase chain reaction (PCR) using marker sequence-specific primers. With this method, both of the clones were assigned to 2q35. These two RFLP markers must be useful for linkage analysis of genetic diseases whose loci are at around 2q35.


Journal of Human Genetics | 1989

Micro extraction of DNA from whole blood and amniocytes

Tetsuya Hirota; Tatsuro Kondoh; Tadashi Matsumoto; Yoshihiro Jinno; Norio Niikawa

SummaryWe describe methods for extracting genomic DNA from a small amount of whole blood or cultured amniocytes. Nuclear DNA was extracted from whole blood spotted on blotting paper. Relatively large molecules of DNA with the average amount of 7–9 μg was extracted from 1 ml of blood spotted and stored for at most two years, being roughly 1/3 of that extracted directly from fresh whole blood. The estimated minimum amount of whole blood that gives a suitable autoradiogram of Southern hybridization was 0.3 ml. Another series of amounts of whole blood or an amniocyte suspension were molded in low-melting agarose into an 100 μl gel block. The DNA extracted from a block that was made from at least 0.25 ml of whole blood, or from 1.25×105 amniocytes (equivalent to 1/8 of the number of confluent cells in a 25 cm2 culture flask) resulted in one suitable Southern analysis. Both methods described here are applicable to the diagnosis of newborns and/or fetuses at risk of a genetic disease and to the diagnosis of a patient from whom a large amount of blood material is difficult to obtain. These methods also make a long-way transportation of the materials possible.


Genomics | 1992

Microdissection of human chromosomal regions 8q23.3–q24.11 and 2q33-qter: Construction of DNA libraries and isolation of their clones

Tetsuya Hirota; Kazuhiro Tsukamoto; Han Xiang Deng; Koh-ichiro Yoshiura; Tohru Ohta; Takaya Tohma; Tetsuya Kibe; Naoki Harada; Yoshihiro Jinno; Norio Niikawa


Journal of Biochemistry | 1992

A simple and efficient amplification method of DNA with unknown sequences and its application to microdissection/microcloning.

Yoshlhlro Jinno; Naoki Harada; Koh-ichiro Yoshiura; Tohru Ohta; Takaya Tohma; Tetsuya Hirota; Kazuhiro Tsukamoto; Han Xiang Deng; Mitsuo Oshimura; Norio Niikawa


American Journal of Medical Genetics Part A | 2003

Very long eyelashes, long eyebrows, sparse hair, and mental retardation in two unrelated boys: An atypical form of Oliver-McFarlane syndrome without retinal degeneration, or a new clinical entity?

Tatsuro Kondoh; Nagisa Amamoto; Tetsuya Hirota; Eiichi Kinoshita; Hiroyuki Moriuchi; Tadashi Matsumoto; Masayuki Shimono; Akihiro Kawakami; Akira Shirahata


Clinical Pediatric Endocrinology | 2003

53 PITUITARY MAGNETIC RESONANCE IMAGING IN CHILDREN WITH GROWTH HORMONE DEFICIENCY

T Shimizu; Katsuaki Motomura; Tomoko Kawaguchi; Eiichi Kinoshita; Tetsuya Hirota; Tsuneyoshi Baba; Masaaki Yoshimoto


Clinical Pediatric Endocrinology | 2002

101 A female case of classical 3β-hydroxysteroid dehydrogenase deficiency with multicystic ovary and anovulatory menstruation : Evaluation of ovarian function after puberty

T Shimizu; Katsuaki Motomura; Tetsuya Hirota; Tomoko Kawaguchi; Eiichi Kinoshita; Tsuneyoshi Baba; Masaaki Yoshimoto


Clinical Pediatric Endocrinology | 2002

98 A male-raised 46, XX case of congenital adrenal hyperplasia due to 21 hydroxylase deficiency

Eiichi Kinoshita; Katsuaki Motomura; T Shimizu; Tetsuya Hirota; Tsuneyoshi Baba; Masaaki Yoshimoto

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Norio Niikawa

Health Sciences University of Hokkaido

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Takaya Tohma

University of the Ryukyus

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