• Title/Summary/Keyword: Chromosomal technology

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Investigation of Transformation Efficiency of Rice Using Agrobacterium tumefaciens and High Transformation of GPAT (glycerol-3-phosphate acyltransferase) Gene Relative to Chilling Tolerance (Agrobacterium tumefaciens를 이용한 벼의 형질전환 효율의 검토 및 내한성 관련 GPAT (glycerol-3-phosphate acyltransferase) 유전자의 형질전환)

  • Seo, Mi-Suk;Bae, Chang-Hyu;Choi, Dae-Ock;Rhim, Seong-Lyul;Seo, Suk-Chul;Song, Pill-Soon;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.29 no.2
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    • pp.85-92
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    • 2002
  • This study has been focused on improving transformation efficiency of rice using Agrobacterium tumefaciens. We have demonstrated the effect of this system when the GPAT gene related to the cold-resistance was transferred by Agrobacterium tumefaciens in rice. Transformation conditions were modified using intron $\beta$-glucuronidase (GUS) expression as a reporter gene in the rice. In this study, mature seed-derived calli of rice (Oruza sativa L. cv. Dongjin) were pre-cultured for 3 days and then infected with Agrobacterium. When this infected calli were cultured in the dark for 10 days on co-cu]lure medium containing 50 mg/L of CaCl$_2$, 30 mg/L of acetosyringone, 2 mg/L of 2,4-D, 120 mg/L of betaine, high GUS expression was observed. In the present transformation system, the efficiency of transformation of GPAT gene was about 54%. Stable integration of GPAT gene into chromosomal DNA was proven by southern blot analysis of genomic DNA isolated from T$_{0}$ progenies. The progenies (T1 generation) derived from primary transformant of 5 lines were segregated with a 3 (resistant) : 1 (sensitive ratio) in medium containing hygromycin. This high frequency transformation system can be used as a useful tool in transformation of another monocotyledon.n.

Analysis of Sequence Type and Fluoroquinolone Resistance in Ciprofloxacin-Resistant Escherichia coli (Ciprofloxacin 내성 대장균에서 Sequence Type과 Fluoroquinolone 내성의 분석)

  • Cho, Hye Hyun
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.3
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    • pp.217-224
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    • 2021
  • Fluoroquinolone (FQ) resistant gram-negative pathogens have emerged worldwide, and the recent increase in FQ resistant Escherichia coli is of great concern in Korea. This study investigated FQ resistance determinants and the epidemiological relationship of 56 ciprofloxacin-resistant E. coli isolated from a tertiary hospital in Daejeon, South Korea from June to December 2018. Molecular epidemiology was investigated by multilocus sequence typing (MLST). Polymerase chain reaction (PCR) and sequence analysis were performed to identify chromosomal mutations in the quinolone resistance determining regions (QRDR) of gyrA, gyrB, parC, and parE and to describe the occurrence of the following plasmid-mediated quinolone resistance (PMQR) genes: aac(6)-Ib-cr, qepA, qnrA, qnrB, qnrC, qnrD, and qnrS. MLST analysis showed 12 sequence types (STs) and the most prevalent ST was ST131 (31/56, 55.4%), followed by ST1193 (13/56, 23.2%), and ST405 (3/56, 5.4%). In 56 ciprofloxacin-resistant E. coli isolates, Ser83→Leu and Asp87→Asn in gyrA and Ser80→Ile and Glu84→Val in parC (51.8%, 29/56) were the most frequent amino acid substitutions and aac(6)-Ib-cr (33.9%, 19/56) was the most common PMQR gene. These results of FQ resistance determinants were more frequently observed in ST131 compared with other clones. Continuous monitoring of the epidemiological characteristics of ciprofloxacin-resistant E. coli isolates and further investigation of FQ resistance determinants are necessary.

Chromosomal Assembly of Tegillarca granosa Genome using Third-generation DNA Sequencing and Hi-C Technology (3세대 DNA 염기서열 분석과 Hi-C기술을 이용한 꼬막 게놈의 유전체 연구)

  • Kim, Jinmu;Lee, Seung Jae;Jo, Euna;Choi, Eunkyung;Cho, Minjoo;Shin, So Ryung;Lee, Jung Sick;Park, Hyun
    • Journal of Marine Life Science
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    • v.6 no.2
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    • pp.97-105
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    • 2021
  • Tegillarca granosa, is one of the most important fishery resources throughout Asia. However, due to industrialization factories, marine environmental pollution, and global warming, the marine fishery production has drop sharply. In order to understand the genetic factors of the blood clam, which is a major fishery resource on the southern coast of Korea, the whole genome of blood clam was studied. The assembled genome of T. granosa was 915.4 Mb, and 19 chromosomes were identified. 25,134 genes were identified, and 22,745 genes were functionally annotated. As a result of performing gene gain and loss analysis between the blood clam genome and eight other types of shellfish, it was confirmed that 725 gene groups were expanded, and 479 gene groups were contracted. The homeobox gene cluster of blood clam showed a well-preserved genetic structure within lophotrochozoan ancestor. T. granosa genome showed high similarity between three hemoglobin genes with Scarpharca broughtonii. The blood clam genome will provide information for the genetic and physiological characteristics of blood clam adaptation, evolution, and the development of aquaculture industry.

G-, C-, and NOR-banding of Korean Native Pig Chromosomes (한국재래돼지의 G-, C-, 및 NOR-banding)

  • Sohn, S.H.;Kweon, O.S.;Baik, K.H.;Jung, W.;Cho, E.J.;Kang, M.Y.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.901-910
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    • 2003
  • Using the G-, C-, and NOR-banding techniques, a karyotyping for Korean Native Pig was performed. Blood samples were collected from 50 male Korean Native Pigs that had been bred at the National Livestock Research Institute and then blood cells were prepared from in vitro cultures followed by karyotyping; G-, C-, and NOR-banding patterns of metaphase chromosomes were analyzed. The karyotype of Korean Native Pig is 38, XX or XY which consists of 5 pairs of submetacentric chromosomes(Group I), 2 pairs of acrocentric chromosomes with short p-arm(Group II), 5 pairs of medium metacentric chromosomes(Group III), 6 pairs of acrocentric chromosomes(Group IV) and metacentric X and Y sex chromosomes. On GTG-banding, the Korean Native Pig exhibited a typical and identical banding pattern in each homologous chromosomes. Overall chromosomal morphology and positions of typical landmarks of the Korean Native Pig were virtually identical to those of Committee for the Standardized Karyotype of the Domestic Pig(CSKDP). However, numbers of G-bands of the Korean Native Pig chromosomes were more than those of CSKDP. In chromosomes 1, 3, 5, 6, 7, 8, 13, 14, 15, 16, 17, 18 and X, the Korean Native Pig exhibited more separated bands as compared with CSKDP. In C-banding patterns, although the quantity of heterochromatin was variable in each chromosome, most of the Korean Native Pig chromosomes had heterochromatic C-bands on centromeres. However, the heterochromatic C-band was constantly observed on the whole Y chromosome. In AgNOR staining, the NORs were located at centromeres on the chromosomes 8 and 10. The number of NORs per metaphase ranged from 2 to 4 giving a mean value of 2.13. The number of NORs were distributed on all chromosome pair 10 but not on chromosome 8. The sizes of NORs were also differed between homologous chromosomes 8. Numbers of NORs of Korean Native Pig were significantly higher than those of Yorkshire. The pattern of pig NORs was polymorphic in breeds, individuals and cells, especially on chromosome 8.

Skewed Inheritance of EST-SSR Alleles in Reciprocal Crosses of Cut Roses (절화장미 품종간 정역교배에 있어서 EST-SSR 마커의 유전)

  • Kim, Jin-Ki;Ahn, Dong-Chun;Oh, Hye-Jeong;Kim, Kwang-Hwan;Choi, Young-Mi;Oh, Seung-Yong;Kang, Nam-Jun;Jeong, Byoung-Ryong;Kim, Zhoo-Hyeon;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.28 no.4
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    • pp.618-626
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    • 2010
  • Matroclinal inheritance of morphological characters in interspecific crosses of Rosa spp. can be influenced by cytoplasmic inheritance, apomixis, and asynaptic heterogamy. In asynaptic heterogamy, which is often observed from interspecific crosses of Rosa sect. $Caninae$, the polyploidy of the seed parent (especially for 5x=35) is recovered in the progeny through the pollens that include only a set of bivalents (x=7) and egg cells that contain a set of bivalents (x=7) and other univalents (3x=21). In this study, we investigated the causes of matroclinal offsprings observed from reciprocal crosses of tetraploid cut rose cultivars ($Rosa$ $hybrida$ L.) by analyzing EST-SSR marker distribution in the progeny populations. From EST-SSR marker analysis of eight offsprings per six reciprocal crosses among six cultivars, cases of cytoplasmic inheritance were not observed. Apomixis was also very rare as compared to the reports on interspecific crosses of sect. $Caninae$; only one apomitic plant was identified from the cross 'Redtem' ${\times}$ 'Red Sandra'. Although a clear-cut pattern of asynaptic heterogamy was not found, cultivar-specific marker transmission skewed to seed parent in four cultivars implied that genetic inheritance can be highly influenced by the seed parent depending on crosses among cut rose cultivars; especially, 10 out of 11 alleles specific to 'Yellow King' distributed in progenies at higher ratios when the cultivars were crossed as the seed parent.

Characterization and Functional Analysis of Obox4 during Oocyte Maturation by RNA Interference (생쥐의 난소와 난자에서의 Obox4의 동정과 RNAi를 이용한 기능연구)

  • Lee, Hyun-Seo;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.4
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    • pp.293-303
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    • 2007
  • Objective: Previously, we identified differentially expressed genes between GV and MII stage mouse oocytes using ACP technology. When we study one of GV selective genes, Obox family, we found Obox4 mRNA expression in ovaries that has been reported as expressed exclusively in testis. Therefore, this study was conducted for characterization and functional analysis for Obox4. Methods: Expression of Obox4 mRNA was examined in gonads and oocytes by RT-PCR. To determine the role of Obox4 in oocyte maturation, Obox4 dsRNA was microinjected into the cytoplasm of GV oocytes followed by 16 h of incubation in the plain medium or by 24 h of incubation in the medium containing IBMX. After RNAi, phenotypes and maturation rates were observed, change in mRNA expression was evaluated, and chromosomal status was confirmed by orcein staining. Results: Obox4 has minimal expression in the ovary compared to that of the other family members. When oocytes were cultured for 16 h in M16 medium after RNAi, maturation rate was not changed significantly, compared with that of non-injected or buffer-injected control oocytes. Surprisingly, however, when oocytes were cultured for 24 h in M16 containing IBMX, in which oocytes were supposed to arrest at GV stage, Obox4 RNAi oocytes were advanced to MI and MII. Spindle structure was disappeared and the chromosomes were condensed in the oocytes after Obox4 RNAi. Conclusions: This is the first report on the expression of Obox4 in the ovary and oocytes. Results of the study suggest that Obox4 plays a crucial role in spindle formation and chromosome segregation during meiosis in oocytes. In addition, Obox4 may play an important role in cAMP-dependent signal cascades of GV-arrest in mouse oocytes.

The Studies on In Vivo Embryo Production using Sex-Sorted Sperm in Korean Brindle Cattle (성 감별 정자를 이용한 칡소 체내수정란 생산에 관한 연구)

  • Jeon, Hyang-A;Moon, Seung-Ju;Lee, Ji-Woong;Kang, Man-Jong;Sohn, Sea-Hwan;Kim, Nam-Tae;Ko, Yeoung-Gyu;Kim, Sung-Woo;Kim, Dong-Kyo;Seong, Hwan-Hoo;Cho, Young Moo;Choe, Changyong
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.47-52
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    • 2016
  • The objective of this study was to investigate the result of in vivo embryo collection and pregnancy rate after embryo transfer using sex-sorted sperm of Korean brindle cattle. Donor Korean brindle cattle superovulation treated by decreasing dose of FSH injection. Embryos were recovered on 7 days after the third artificial insemination. Control group semen straw used artificial insemination contained 20 million sperm. Sex-sorted semen straws contained 4 million sperm or 10 million sperm. As for the result of the recovery of the in vivo embryos derived from sex-sorted sperm, the number of transferable embryos was significantly highly recovered to be $6.20{\pm}2.28/donor$ from the control group and was significantly lowly recovered to be $1.57{\pm}1.72/donor$ from the group treated at a sperm concentration of $10{\times}10^6$ (p<0.05). The number of unfertilized embryo was $0.8{\pm}1.30/donor$ in control group which was significantly lower than the group treated at a sperm concentration of $4{\times}10^6$ (p<0.05). However, there was no significant difference in the number of undeveloped ova between control and treatment groups. Pregnancy rate after embryo transfer was shown to be 35.00% in control group and 12.50% in treatment group. The karyotype analysis of the calf derived from sex-sorted sperm resulted in a similar chromosomal distribution pattern (2n=60, XX) compared to those of common Korean native cattle.

Down syndrome in women aged more than 35 Years positive detection rates (산전선별검사를 통한 35세이상 산모 다운증후군 양성률 비교 평가)

  • Oh, Taek Min;Kim, Ga-Yeon;Lee, Young ki
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.22 no.6
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    • pp.314-320
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    • 2021
  • With the increasing age of motherhood in recent years, attributed to late marriages due to social or environmental factors, the Down's syndrome screening test using biochemical markers has become essential for pregnant women. The process of diagnosing Down's syndrome pregnancy in the high-risk group subjects involves chromosomal analysis, which is performed on samples obtained through invasive procedures such as chorionic biopsy or amniotic fluid. Thus, to reduce unnecessary invasive tests and lower the risk to mother and fetus, it is important to identify a screening test with low risk and high Down's syndrome detection rate. Recently, as the average age of mothers has increased, numerous inspection agencies have classified high-risk mothers as women over the age of 35 years. This study evaluated a total of 36,436 pregnant women aged between 17 to 46 years, and who requested prenatal screening at an inspection agency in Yongin in 2018. Test (13,690 people) Four tests were conducted by applying the time-resolved fluoroimmunoassay method using the direct sandwich and indirect sandwich technology, and the immunoassay method using the sandwich method. We aimed to confirm the difference in positivity rate with increasing age of the subjects. We believe that in future, data obtained from this study will be very useful for the prevention and treatment of Down's syndrome risk at varied inspection institutions, and for prospective mothers.

A Study of Guidelines for Genetic Counseling in Preimplantation Genetic Diagnosis (PGD) (착상전 유전진단을 위한 유전상담 현황과 지침개발을 위한 기초 연구)

  • Kim, Min-Jee;Lee, Hyoung-Song;Kang, Inn-Soo;Jeong, Seon-Yong;Kim, Hyon-J.
    • Journal of Genetic Medicine
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    • v.7 no.2
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    • pp.125-132
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    • 2010
  • Purpose: Preimplantation genetic diagnosis (PGD), also known as embryo screening, is a pre-pregnancy technique used to identify genetic defects in embryos created through in vitro fertilization. PGD is considered a means of prenatal diagnosis of genetic abnormalities. PGD is used when one or both genetic parents has a known genetic abnormality; testing is performed on an embryo to determine if it also carries the genetic abnormality. The main advantage of PGD is the avoidance of selective pregnancy termination as it imparts a high likelihood that the baby will be free of the disease under consideration. The application of PGD to genetic practices, reproductive medicine, and genetic counseling is becoming the key component of fertility practice because of the need to develop a custom PGD design for each couple. Materials and Methods: In this study, a survey on the contents of genetic counseling in PGD was carried out via direct contact or e-mail with the patients and specialists who had experienced PGD during the three months from February to April 2010. Results: A total of 91 persons including 60 patients, 49 of whom had a chromosomal disorder and 11 of whom had a single gene disorder, and 31 PGD specialists responded to the survey. Analysis of the survey results revealed that all respondents were well aware of the importance of genetic counseling in all steps of PGD including planning, operation, and follow-up. The patient group responded that the possibility of unexpected results (51.7%), genetic risk assessment and recurrence risk (46.7%), the reproduction options (46.7%), the procedure and limitation of PGD (43.3%) and the information of PGD technology (35.0%) should be included as a genetic counseling information. In detail, 51.7% of patients wanted to be counseled for the possibility of unexpected results and the recurrence risk, while 46.7% wanted to know their reproduction options (46.7%). Approximately 96.7% of specialists replied that a non-M.D. genetic counselor is necessary for effective and systematic genetic counseling in PGD because it is difficult for physicians to offer satisfying information to patients due to lack of counseling time and specific knowledge of the disorders. Conclusions: The information from the survey provides important insight into the overall present situation of genetic counseling for PGD in Korea. The survey results demonstrated that there is a general awareness that genetic counseling is essential for PGD, suggesting that appropriate genetic counseling may play a important role in the success of PGD. The establishment of genetic counseling guidelines for PGD may contribute to better planning and management strategies for PGD.