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Identification of Compound Heterozygous Alleles in a Patient with Autosomal Recessive Limb-Girdle Muscular Dystrophy (상염색체 열성 지대형 근이영양증 환자로부터 TTN 유전자의 복합 이형접합성 대립유전자의 분리)

  • Choi, Hee Ji;Lee, Soo Bin;Kwon, Hye Mi;Choi, Byung-Ok;Chung, Ki Wha
    • Journal of Life Science
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    • v.31 no.10
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    • pp.913-921
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    • 2021
  • Limb-girdle muscular dystrophy (LGMD) which is characterized by progressive muscle weakening of the hip and shoulder shows both dominant and recessive inheritances with many pathogenic genes including TTN. This study performed to identify genetic causes of a male patient with late onset (45 years old) autosomal recessive LGMD and atrial flutter. By application of the whole exome sequencing, we identified bi-allelic variants of TTN gene in the patient. One allele had a single missense variant of [c.24124G>T (p.V8042F)], while the other allele consisted of three missense variants of [c.29222G>C (p.R9741P) + c.67490A>G (p.H22497R) + c.75376C>T (p.R25126C)]. The p.V8042F allele was transmitted from his mother, while the other haplotype allele was putatively transmitted from his father. His two unaffected sons had only the p.R9741P. These variants have been not reported or rarely reported in the public human genome databases (1,000 Genome, gnomAD, and KRGDB). Most variants were located in the highly conserved immunoglobulin or fibronectin domains and were predicted to be pathogenic by the in silico analyses. The TTN giant protein plays a key role in muscle assembly, force transmission at the Z-line, and maintenance of resting tension in the I-band. In conclusion, we think that these bi-allelic compound heterozygous mutations may play a role as the genetic causes of the LGMD phenotype.

Anti-inflammatory Activities of Antimicrobial Peptide Locustacin Derived from Locusta migratoria in LPS-stimulated RAW264.7 Cells (풀무치 유래 항균 펩타이드 locustacin의 항염증 활성)

  • Choi, Ra-Yeong;Lee, Joon Ha;Seo, Minchul;Kim, In-Woo;Hwang, Jae-Sam;Kim, Mi-Ae
    • Journal of Life Science
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    • v.31 no.10
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    • pp.898-904
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    • 2021
  • Locusta migratoria is a widespread locust species in many parts of the world and is considered an alternative source for the production of protein for value-added ingredients. We previously identified putative antimicrobial peptides derived from L. migratoria through an in silico analysis of its transcriptome. However, its anti-inflammatory effect has not been studied. In this study, we investigated the anti-inflammatory activities of the antimicrobial peptide locustacin (KTHILSFFPSFLPLFLKK-NH2) derived from L. migratoria on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Locustacin (50, 100, and 200 ㎍/ml) significantly reduced the production of nitric oxide (NO) in LPS-stimulated macrophages without any cytotoxicity. Locustacin also inhibited the mRNA and protein expression of pro-inflammatory mediators, such as inducible NO synthase and cyclooxygenase-2, in contrast to the presence of LPS alone. Locustacin decreased the release of LPS-induced pro-inflammatory cytokines, including interleukin (IL)-6 and IL-1β, and their gene expression in a dose-dependent manner. Furthermore, locustacin (100 and/or 200 ㎍/ml) inhibited phosphorylation levels of extracellular signal regulated kinase, p38, and c-Jun N-terminal kinase. Locustacin also suppressed the degradation of inhibitory kappa B alpha, which was considered to be an inhibitor of nuclear factor kappa B (NF-κB). Collectively, these results demonstrate that locustacin can exert anti-inflammatory effects through the inhibition of mitogen-activated protein kinase (MAPK) phosphorylation, activation of NF-κB, and downstream inflammatory mediators in LPS-stimulated macrophage cells.

Thrips Infesting Hot Pepper Cultured in Greenhouses and Variation in Gene Sequences Encoded in TSWV (시설재배지 고추를 가해하는 총채벌레류와 TSWV 유전자 서열 변이)

  • Kim, Chulyoung;Choi, Duyeol;Kang, Jeong Hun;Ahmed, Shabbir;Kil, Eui-Joon;Kwon, Gimyeon;Lee, Gwan-Seok;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.60 no.4
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    • pp.387-401
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    • 2021
  • Thrips infesting hot peppers were monitored in greenhouses using yellow sticky traps. In addition, the hot peppers infected with tomato spotted wilt virus (TSWV) were observed during the monitoring period. The flower thrips (Frankliniella intonsa) were initially trapped at a low density just after transplanting seedlings of hot peppers at late March. The western flower thrips (Frankliniella occidentalis) were trapped after mid April. These two thrips represented more than 98% of the total thrips attracted to the traps after May, in which F. intonsa showed higher occurrence frequency than F. occidentalis. The total number of thrips had two peaks at mid May with a small and short-term peak and at June-July with a large and long-term peak. The trapped thrips exhibited inconsistent sex ratios, suggesting a seasonal parthenogenesis. Different geographical populations were varied in cytochrome oxidase I sequences, in which local populations in Andong shared a high sequence similarity. TSWV-infected hot peppers, which might be mediated by these two thrips species, were observed and confirmed by an immunoassay kit and a molecular diagnosis using RT-PCR. In addition, the TSWV was detected in F. occidentalis collected from the infected hot peppers. Three open reading frames (NSS, N, and NSM) of the isolated TSWV genomes were sequenced and showed multiple point mutations containing missense mutations among geographical variants. When the isolated TSWV was fed to nonvirulent thrips of F. occidentalis, the virus was detected in both larvae and adults. However, the viral replication occurred in larvae, but not in adults.

Hair growth promoting effect of toothpaste in C57BL/6 mice: Active components and their effects on genomic expression (C57BL/6 마우스에서 치약의 모발성장 촉진 효과: 유효 성분과 유전체 발현에 미치는 영향)

  • Ahn, Seunghyun;Lee, Jung Yeon;Shin, Yujeong;Lee, Jinkyung;Lee, Seol-Hoon;Park, Seyeon
    • Journal of Applied Biological Chemistry
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    • v.64 no.4
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    • pp.421-431
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    • 2021
  • It has been suggested that some toothpastes have the potential to promote hair growth. However, there was no scientific verification on the hair growth effect of toothpaste and no scientific report on major active ingredients in toothpaste. In this work, toothpaste and its constituents were applied topically over the shaved skin of C57BL/6 mice and evaluated. Results indicated that toothpaste showed hair growth effect. Also, the effect of toothpaste constituents on the proliferation rate of keratinocyte cells was investigated. The mixture solution of 𝛼-tocopherol acetate, l-menthol, and stevioside, each of that was known to promote hair growth and other toothpaste constituents were applied topically on mouse skin. When the mixture solution was included, hair growth effect was observed in mice. Transcriptome analysis was performed using the dorsal epidermis of mice from the group treated with toothpaste, the mixture which are presumed to be active ingredients for hair growth, and from mice used for the control group. As a result of analyzing the genes whose expression was significantly changed in each treatment group, the gene patterns of the two groups were very similar. Also, when functional genomic analysis was performed, genes with functions related to hair growth regulation showed a high extent of the change in both groups. Hair growth-related genes whose expression was changed in both groups included keratin, keratin-related proteins, forkhead box, and sonic hedgehog. Therefore, the hair growth effect of toothpaste is thought to be due to the effect of a mixture of 𝛼-tocopherol acetate, l-menthol, and stevioside.

Association Analysis of the 6-bp Deletion Variant of the MYH3 Gene with Meat Color Traits in Crossbred (Landrace × Jeju Native Black Pig) Pigs (제주재래흑돼지와 랜드레이스 교배 축군에서 MYH3 유전자의 6-bp 결실 변이와 육색 형질간의 연관성 분석)

  • Kang, Yong-Jun;Kim, Sang-Geum;Kim, Su-Yeon;Kim, Min-Ji;Kim, Hyeon-Ah;Shin, Moon-Cheol;Yoo, Ji-Hyun;Yang, Byoung-Chul;Cho, In-Cheol
    • Journal of Life Science
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    • v.31 no.7
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    • pp.626-630
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    • 2021
  • This study was conducted to examine the association between the myosin heavy chain 3 (MYH3) in 6-bp deletion variant genotypes and meat color traits in a crossbred pig population Landrace and Jeju native black pigs (JNBP). The longissimus dorsi, semimembranosus, triceps brachii and biceps femoris muscle from each carcass were used for the analysis of meat color traits. A total of 187 pigs and three meat color traits, CIE L* (lightness), CIE a* (redness), and CIE b* (yellowness), were analyzed. All experimental pigs were successfully genotyped for the MYH3 6-bp deletion variant using Polymerase chain reaction (PCR) analysis. We detected three MYH3 6-bp deletion variant genotypes qq, Qq, and QQ with 0.091, 0.551 and 0.358 genotype frequencies, respectively. Compared to qq homozygotes, the MYH3 6-bp deletion QQ genotype animals showed a higher levels of the meat colors traits CIE L* (lightness), CIE a* (redness), and CIE b* (yellowness) in longissimus dorsi (p>0.05, p<0.001, p<0.001), semimembranosus (p>0.05, p<0.001, p<0.001), triceps brachii (p<0.001, p<0.001, p<0.001), and biceps femoris (p<0.01, p<0.001, p<0.001), respectively. The QQ genotype pigs was associated with increasing meat color traits in the crossbred between Landrace and JNBP. Our findings suggest that the MYH3 6-bp deletion variant genotypes can be used as valuable genetic markers for JNBP-related breeding programs to improve meat quality and control meat color traits.

Effect of 17β-estradiol on Ecdysteroid Pathway Related Genes in the Brackish Water Flea Diaphanosoma celebensis (17β-estradiol이 기수산 물벼룩의 Ecdysteroid 경로에 미치는 영향)

  • In, Soyeon;Yoo, Jewon;Cho, Hayoung;Lee, Young-Mi
    • Journal of Marine Life Science
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    • v.5 no.2
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    • pp.35-42
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    • 2020
  • 17β-estradiol (E2) is a natural hormone secreted by ovary, and continuously discharged from household and livestock wastewater into aquatic environment. Due to its strong estrogenic activity, it has adverse effects on development and reproduction in crustacean as an endocrine disrupting chemical. Although ecdysteroid signaling pathway play a key role in development in crustacean, little information on transcriptional modulation of ecdysteroid-related genes in response to E2 is available in small crustacean. Here, we investigated the acute toxicity of E2 to obtain 24-h LCx values in the brackish water flea Diaphanosoma celebensis. Time-dependent expression patterns of seven ecdysteroid pathway - related genes (CYP314a1, EcRA, EcRB, USP, ERR, Vtg, VtgR) were further examined using quantitative real time reverse transcriptase polymerase chain reaction (qRT-PCR). As results, 24-h LC50 and LC10 values were 9.581 mg/l and 4.842 mg/l, respectively. The mRNA expression of CYP314a1, EcRA, USP, VtgR was significantly up-regulated at 12 or 24 h after exposure to E2. These findings indicate that E2 can affect their molting and reproduction by modulating the expression of ecdysteroid pathway - related in D. celebensis. This study will be useful for better understanding of molecular mode of action of endocrine disrupting chemicals on molting process in small crustacean.

Assessment Selective Breeding Effect of Israeli carp (Cyprinus carpio) from Korea (국내 이스라엘 잉어의 선발육종효과 평가)

  • Kim, Jung Eun;Hwang, Ju-ae;Kim, Hyeong Su;Im, Jae Hyun;Lee, Jeong-Ho
    • Korean Journal of Ichthyology
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    • v.32 no.4
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    • pp.210-221
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    • 2020
  • Since the introduction of Israeli carp into Korea for farming in 1973, there are no breeding studies on developing Korea Israeli carp (domestic) so far. This study performed gene-based cross-breeding studies to restore genetic diversity of lowered Israeli carp through continuous inbreeding, and for rapid growth and better scales. This study produced four cross-breeding groups (F1) using Koean Israeli carp and Chinese Songpu mirror carp for the improvement of growth and scale of Israeli carp in Korea. And mating scheme for breeding groups was set in consideration of the morphological analysis and genetic distance of broodstock. In addition, this study used microsatellite markers and genotype data to analyze genetic diversity and parentage analysis. As a result, the average NA and HE values of Korean select broodstock are 8.3 and 0.743, and F1 is 13.0 and 0.764. This study shows that the genetic diversity of F1 has been recovered over Korean Israeli carp through breeding between Korean Israeli carp and Chinese Songpu mirror carp. Common Israeli carp in Korea reached 1.7 kg in 17 months, and improved Israeli carp reached to 2.2 kg. The KC (Korea×China, KC) group was 2.52 and broodstock group was 3.15. F1 showed lower scale score (0.63) than broodstock. The improved carp (F1; CK, KC) had 20% better scales than the parent group (F0), which improved 27% in weight and 25% in scales compared to common Israeli carp. The Israeli carp developed by the genetics-based breeding grew quicker and had improved genetic diversity and fewer scales, which will be of great value for Korean Israeli aquaculture industry due to good marketability.

Bacterial Blight Resistant Mid-late Maturing Rice 'Manbaek' with High Grain Quality (벼흰잎마름병 저항성 고품질 중만생 벼 '만백')

  • Park, Hyun-Su;Baek, Man-Kee;Kim, Bo-Kyeong;Kim, Ki-Young;Shin, Woon-Chul;Ko, Jae-Kwon;Nam, Jeong-Kwon;Kim, Woo-Jae;Cho, Young-Chan;Ko, Jong-Cheol;Kim, Jeong-Ju;Kim, Hyun-Soon
    • Korean Journal of Breeding Science
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    • v.49 no.3
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    • pp.235-244
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    • 2017
  • 'Manbaek' is a bacterial blight resistant mid-late maturing rice cultivar with high grain quality. 'Manbeak' was derived from anther culture using the backcross combination, $Hopum^*2/SR30075$. 'Hopum' is a mid-late maturing rice cultivar with high grain quality and 'SR30075' is a pyramid line carrying three bacterial blight resistance genes. 'Manbaek' was selected through the pedigree method, yield trials, and local adaptability tests. 'Manbeak' carrying two bacterial blight resistance gene Xa3 and xa5 showed high-level and broad-spectrum resistance against bacterial blight. 'Manbaek' was resistant to K3a, mostly virulent race in Korea, and exhibited resistance reaction against 16 Korean bacterial blight isolates. 'Manbaek' was a mid-late maturing rice. The heading date of 'Manbaek' was August 19th, which was 5 days later than that of 'Nampyeong'. Manbaek' was a lodging-tolerant rice with short culm and dark green leaf. Due to the low viviparous germination, 'Manbaek' could be a useful material to prevent pre-harvest sprouting. 'Mabeak' was resistant to bacterial blight and rice stripe virus, but susceptible to other virus diseases and insect pests. The yield of 'Manbaek' was similar to 'Nampyeong'. 'Manbaek' showed excellent grain appearance and good tastes of cooked rice, so that it could contribute to improving the quality of bacterial blight resistant cultivars. 'Manbaek', bacterial blight resistant cultivar with high grain quality, is suitable for the cultivation at bacterial blight prone area and has been utilized in the breeding programs for enhancing the resistance against bacterial blight (Registration No. 6069).

Genetic Variation of Parental Inbred Lines for Korean Waxy Corn Hybrid Varieties revealed by SSR markers (우리나라 찰옥수수 품종들의 교배친 자식계통들에 대한 유전적 변이성)

  • Park, Jun-Sung;Sa, Kyu-Jin;Park, Ki Jin;Jang, Jin-Sun;Lee, Ju Kyong
    • Korean Journal of Breeding Science
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    • v.41 no.2
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    • pp.106-114
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    • 2009
  • In maize, knowledge of genetic diversity and genetic relationships among elite inbred lines is an significant impact on the selection of parental lines for hybrid varieties. Genetic diversity and genetic relationships among 11 parental inbred lines of Korean waxy and normal corn varieties were analyzed using 50 SSR markers distributed over the whole genome. A total of 171 allele bands were detected with an average of 3.4 alleles per locus. Number of allele bands per locus ranged from two to six and gene diversity varied from 0.165 to 0.900 with an average of 0.596 depending on the SSR loci. The cluster tree recognized three major groups with 61.6% genetic similarity. Group I includes 7 inbred lines (KL103, HW1, HW4, HW6, HW7, HW8, HW9), with similarity coefficients of between 0.616 and 0.730. Group II includes 2 inbred lines (HF1, HF2), with similarity coefficients of 0.959. Group III includes 2 inbred lines (HW3, HW5), with similarity coefficients of 0.713. The present study indicates that the SSR markers chosen for this analysis are effective for the assessment of genetic diversity and genetic relationships among 11 parental inbred lines.

Identification of Leaf Blast Resistance Genes Derived from a Korean Weedy Rice, Ganghwaaengmi 11 (잡초성벼인 강화앵미11 유래 잎도열병 저항성 유전자 탐색)

  • Suh, Jung-Pil;Cho, Young-Chan;Kim, Jeong-Ju;Shin, Young-Seop;Yang, Chang-Ihn;Roh, Jae-Hwan;Kim, Yeon-Gyu
    • Korean Journal of Breeding Science
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    • v.42 no.4
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    • pp.390-396
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    • 2010
  • A weedy rice, Ganghwaaengmi 11, shows high level of leaf blast resistance. The chromosomal number and locations of genes conferring the leaf blast resistance were detected by QTL (quantitative trait loci) analysis using SSR markers in the 120 RILs (recombinant inbred lines) derived from the cross between Nagdongbyeo and Ganghwaaengmi 11. Ganghwaaengmi 11 expressed compatibility with 20 of the 45 inoculated blast isolates, in contrast to Nagdongbyeo with 44 compatible isolates. To identify QTLs affecting partial resistance, RILs were assessed in upland blast nursery in three regions and inoculated with selected nine blast isolates. QTLs for resistance to blast isolates were identified on chromosomes 7, 11 and 12. Three QTLs associated with blast resistance in nursery test at three regions were also detected on chromosomes 7, 11 and 12. The QTL commonly detected on chromosome 12 was only increased blast resistance by Ganghwaaengmi 11 allele. This QTL accounted for 60.3~78.6% of the phenotypic variation in the blast nursery test. OSR32 and RM101 markers tightly linked to QTL for blast resistance on chromosome 12 might be useful for marker-assisted selection (MAS) and gene pyramiding to improve the blast resistance of japonica rice.