• Title/Summary/Keyword: Hybridization, Genetic

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An update of preimplantation genetic diagnosis in gene diseases, chromosomal translocation, and aneuploidy screening

  • Chang, Li-Jung;Chen, Shee-Uan;Tsai, Yi-Yi;Hung, Chia-Cheng;Fang, Mei-Ya;Su, Yi-Ning;Yang, Yu-Shih
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.3
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    • pp.126-134
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    • 2011
  • Preimplantation genetic diagnosis (PGD) is gradually widely used in prevention of gene diseases and chromosomal abnormalities. Much improvement has been achieved in biopsy technique and molecular diagnosis. Blastocyst biopsy can increase diagnostic accuracy and reduce allele dropout. It is cost-effective and currently plays an important role. Whole genome amplification permits subsequent individual detection of multiple gene loci and screening all 23 pairs of chromosomes. For PGD of chromosomal translocation, fluorescence $in-situ$ hybridization (FISH) is traditionally used, but with technical difficulty. Array comparative genomic hybridization (CGH) can detect translocation and 23 pairs of chromosomes that may replace FISH. Single nucleotide polymorphisms array with haplotyping can further distinguish between normal chromosomes and balanced translocation. PGD may shorten time to conceive and reduce miscarriage for patients with chromosomal translocation. PGD has a potential value for mitochondrial diseases. Preimplantation genetic haplotyping has been applied for unknown mutation sites of single gene disease. Preimplantation genetic screening (PGS) using limited FISH probes in the cleavage-stage embryo did not increase live birth rates for patients with advanced maternal age, unexplained recurrent abortions, and repeated implantation failure. Polar body and blastocyst biopsy may circumvent the problem of mosaicism. PGS using blastocyst biopsy and array CGH is encouraging and merit further studies. Cryopreservation of biopsied blastocysts instead of fresh transfer permits sufficient time for transportation and genetic analysis. Cryopreservation of embryos may avoid ovarian hyperstimulation syndrome and possible suboptimal endometrium.

Production and Characterizations of Somatic Hybrids between Brassica campestris L. ssp pekinensis and Brassica of oleracea L. var capitata

  • Lian, Yu-Ji;Lim, Hak-Tae
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.33-38
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    • 2001
  • Protoplasts isolated from inbred lines of Brassica oleracea L. var capitata (cabbage) and Brassica campestris L. ssp. pekinensis (Chinese cabbage) were fused by PEG-mediated method, and somatic hybrid cells were differentiated into plants. for the identification of somatic hybrid plants, ploidy level, plant morphology, and cytological analysis were performed. All of the regenerated plants derived from fused protoplasts were shown to be 2X-4X, or higher ploidy level, presumably due to somatic hybridization or chromosome doubling. The morphology of leaves, petioles, and flowers showed an intermediate phenotype between Chinese cabbage and cabbage. Chromosome numbers in these somatic hybrids ranged mostly from 33 to 38. According to Genomic in situ hybridization (GISH) pattern, signals from both fusion parents of B.campestris or B.oleracea were detected in different colors when chromosomes of putative somatic hybrids were observed.

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Comparative genomic hybridization analysis of fetal chromosomal aberrations

  • Choi, Soo-Kyung;Kim, Young-Mi;Park, So-Yeon;Kim, Jin-Woo;Ryu, Hyun-Mee;Go, Chang-Won;Park, Chong-Tak;Jun, Jung-Young;Park, In-Suh
    • Journal of Genetic Medicine
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    • v.2 no.2
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    • pp.71-77
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    • 1998
  • Comparative genomic hybridization (CGH) can now be applied to detect the origin of extra or missing chromosomal material in cases with common unbalanced aberrations and in prenatal investigations. This method has been used in 13 cases of fetal samples for this study; 3 for amniocytes, 2 for cord blood and 8 for abortus tissues. These samples were previously subjected to GTG-banding. Our study showed aneuploidy in 8 cases, and partial monosomy, partial trisomy or marker chromosome in the remaining 5. The CGH disclosed further small genetic imbalances in 4 of all 13 cases: a prenatal sample showing del(20)(q13) by GTG confirmed a loss of the segment 20p13-pter by CGH; a marker chromosome manifested normal CGH profile; chromosome der(?)(?;15) found in an abortus sample by GTG turned out to be a loss of 15pter-q14 (partial monosomy) and a gain of 10pter-q22 (partial trisomy); the der(15) shown by GTG represented partial trisomy of 3q24-qter. These findings show that CGH is very useful and efficient for cytogenetic investigations of clinical cases.

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Distinctive Characteristics of an Autonomous Replication Sequence of Cephalosporium acremoniurn in Yeast (Cephalosporium acremonium의 자율복제 기점의 특성)

  • Lee, Kyoung;Kang, Dae-Wook;Yoon, Byung-Dae;Hwang, In-Kyu;Ahn, Jong-Seog;Min, Tae-Ick
    • Microbiology and Biotechnology Letters
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    • v.19 no.3
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    • pp.215-221
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    • 1991
  • An autonomous replication sequence (ARS) derived from Cephalosporium acremonium ATCC 20339 was cloned in Sarchuromyces cerevisiae SHY 3 using YIp5 as a cloning vector. A new recombinant plasmid, designated pCY-2, which contained a 3.7 kb BamHI fragment of C. acrenzonium DNA showed the highest stability among the 40 recombinant plasmids composed of the YIp5 2nd ARS of C. ucremoniztm. Also, Southern hybridization and transformation of E, cull with DNA purified from yeast transformants verified that pCY-2 autonomously replicates in yeasts. Transformation efficiency and plasmid stability of pCY-2 in yeast were higher than those ol YRp 7 containing ARS which originated from yeast. Detailed studies by subcloning revealed that two ARSs existed within 2.6 kb of the insert, which is a novel discovery. However, it was concluded that these two ARSs were ligated during the gene manipulation in vitro.

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COMPARATIVE GENOMIC HYBRIDIZATION STUDIES ON CHOLANGIOCARCINOMA IN KOREA

  • Lee, Chul-Hoon;Park, Bu-Young;Kim, Min--Kyoung;Jee- Hong Kyhm;Park, Ho-Soon;Cho, Youl-Hee
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.10a
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    • pp.190-190
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    • 2001
  • The elucidation of the genetic changes of cholangiocarcinoma is very important for understanding the molecular mechanism of carcinogenesis and progression of cholangiocarcinoma. In order to identify the gains or losses of the copy number of DNA sequence in cholangiocarcinoma, we used comparative genomic hybridization to study 33 cases of cholangiocarcinoma. The whole DNAs from each tumor tissue were labeled with different fluorochromes and then simultaneously hybridized to normal metaphase spread chromosomes.(omitted)

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Classification and Genetic Variation Analysis Among Formae Speciales of Fusarium oxysporum by Using Recombinant DNA Probes (재조합 DNA probe에 의한 Fusarium oxysporum 분화형간의 분류 및 유전적 변이 분석)

  • Kim, Young-Tae;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.25 no.4 s.83
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    • pp.362-368
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    • 1997
  • Five formae speciales of Fusarium oxysporum in Korea were examined using RFLP analysis to find the possibility for classification and analyze genetic variations. DNAs from F. oxysporum f. sp. lycopersici, cucumerinum, fragariae, garlic and sesami were used with three recombinant probes such as pFC46, pFC52 and pFC57. Distinct differences among five formae speciales of this fungus were detected in RFLP band patterns based on southern hybridization of genomic DNA using each recombinant clone, which was a repetitive copy probe. Strains belong to four formae speciales could be very stable in genetic variation except f. sp. sesami which has more variation than the others based on the RFLP analysis. They formed their own cluster which has high similarity within the same formae specialis resulted from the UPGMA analysis for genetic relationship analysis and each cluster represented its own formae specialis. The method using three recombinant DNA probes could be a good tool for classification of formae speciales in F. oxysporum.

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Clinical Application of Chromosomal Microarray for Germline Disorders

  • Chang Ahn Seol
    • Journal of Interdisciplinary Genomics
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    • v.5 no.2
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    • pp.24-28
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    • 2023
  • Chromosomal microarray (CMA) is primarily recommended for detecting clinically significant copy number variants (CNVs) in the genetic diagnosis of developmental delay, intellectual disability, autism, and congenital malformations. Prenatal CMA is recommended when a fetus has major congenital malformations. The main principles of CMA can be divided into array comparative genomic hybridization and single-nucleotide polymorphism arrays. In the current CMA platforms, these two principles are combined, and detection of genetic abnormalities including CNVs and absence of heterozygosity is facilitated. In this review, I described practical assessment of CMA testing regarding to laboratory management of CMA, interpretation of CNVs, and special considerations for comprehensive genetic counseling.

Construction of cDNA Library from Posterior Silk Gland (PSG) of Korean Oak Silkmoth, Antheraea yamamai and Molecular Cloning of Fibroin Heavy Chain Gene(FHC)

  • Lee, Jin-Sung;Kim, Soon-Jung;Kim, Ki-Hwan;Park, Young-Min;Suh, Dong-Sang
    • Journal of Life Science
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    • v.10 no.1
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    • pp.10-13
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    • 2000
  • To develope the genetic source of oak wild silkworm, Antheraea yamamai, the cDNA library was constructed with poly A+ mRNA isolated from posterial silk gland of fifth instar larvae. Titer of the cDNA library was about 5.1$\times$105 pfu in total. We presumed that the titer covered almost all transcripts existed in Antherea yamamai. From cDNA library of Antheraea yamamai, fibroin heavy chain gene, which is specifically expressed from posterial silk gland of Antheraea yamamai, was screened using oligonuclotide probe specific to alanine rich motif of fibrin heavy chain gene of Antheraea pernyi. As a result, fibroin clones isolated from 5$\times$104 plaques showed the highest homolgy (95%) with that of Antherea pernyi in nucleotide of Anthereaea yamamai and Bombyx mori shows that there is no homologous sequence in the 3+ partial 채야후 region Genomic southern hybridization suggested that one copy is present. Northern hybridization showed that fibroin transcript was approximateely 9 kb in length.