• Asian-Australasian Journal of Animal Sciences
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• 제31권4호
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• pp.489-498
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• 2018

Objective: Foot and mouth disease (FMD) and porcine reproductive and respiratory syndrome (PRRS) are major diseases that interrupt porcine production. Because they are viral diseases, vaccinations are of only limited effectiveness in preventing outbreaks. To establish an alternative multi-resistant strategy against FMD virus (FMDV) and PRRS virus (PRRSV), the present study introduced two genetic modification techniques to porcine cells. Methods: First, cluster of differentiation 163 (CD163), the PRRSV viral receptor, was edited with the clustered regularly interspaced short palindromic repeats-CRISPR-associated protein 9 technique. The CD163 gene sequences of edited cells and control cells differed. Second, short hairpin RNA (shRNAs) were integrated into the cells. The shRNAs, targeting the 3D gene of FMDV and the open reading frame 7 (ORF7) gene of PRRSV, were transferred into fibroblasts. We also developed an in vitro shRNA verification method with a target gene expression vector. Results: shRNA activity was confirmed in vitro with vectors that expressed the 3D and ORF7 genes in the cells. Cells containing shRNAs showed lower transcript levels than cells with only the expression vectors. The shRNAs were integrated into CD163-edited cells to combine the two techniques, and the viral genes were suppressed in these cells. Conclusion: We established a multi-resistant strategy against viral diseases and an in vitro shRNA verification method.

• 한국미생물·생명공학회지
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• 제43권1호
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• pp.22-30
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• 2015

CSF is a major concern for the swine industry, representing currently the most epizootically dangerous disease to the species. Numerous CSFV isolates with various degrees of virulence have already been isolated worldwide, ranging from low virulent strains that do not result in any apparent clinical signs to highly virulent strains that cause a severe per acute hemorrhagic fever with very high mortality. The molecular epidemiology of CSFVs has proven to be an essential tool for effective disease control and the development of safe and effective vaccines. Therefore, this study cloned and sequenced local CSFV isolates, and conducted a phylogenetic analysis based on the E2 glycoprotein encoding sequences.The RNA was extracted from PK15 cell culture passaged CSFV isolates, the cDNA prepared, and the complete E2 gene amplified with a product size of 1186 bp. The gelpurified PCR product was cloned into a pGEMT easy vector and the positive clone commercially sequenced. Aligning the nucleotide (1119 bp) and amino acid (373) sequences with 29 reference strains revealed nucleotide and amino acid sequence identities of 82.60-97.80% and 88.70-98.70%, respectively, indicating a higher mutation rate of the field CSFV strains. The phylogenetic analysis based on the complete E2 amino acid sequences also revealed a reliable differentiation of all the analyzed strains into specific genetic groups and subgroups, plus the local isolate (CSFV-E2) was found to cluster with the CSFV subgroup 2.2. Thus, the full-length E2 cds proved to be most suitable for a reliable and statistically significant phylogenetic analysis of CSFV isolates.

• Journal of Animal Science and Technology
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• 제47권2호
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• pp.159-166
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• 2005

Porcine myostatin(MS1N) gene plays a key role in the differentiation of myoblast and muscle development. Genetic polymorphism was screened by single stranded conformation polymorphism(SSCP) analysis and subsequent DNA sequencing detected a nucleotide substitution(C2150T) in exon 3 of MSIN gene. Phenotypic association of the polymorphism was tested in a Landrace population and positive effects of the allele T for lean growth traits were found in the population. Even though it is not significant, the pigs have IT and TC genotypes were heavier for the body weight at birth and at twenty weeks of age than those containing genotype. Cc. However, the allele T was significantly associated with higher eye muscle area(P < 0.05). As a result of this study, we suggested that the allele T in exon 3 of MSTN gene comes a significant effect for increasing the eye muscle area without decreasing backfat thickness. This polymorphism did not change the amino acid but Taq I -RFLP matched to SSCP band patterns in exon 3 of MSTN gene, which will be an useful molecular marker for breeding of Landrace pigs.

• Animal Systematics, Evolution and Diversity
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• 제29권2호
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• pp.136-143
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• 2013

The collared scops owl that occurs in Korea is a protected species but its exact specific status has been questioned. To resolve the species status, a molecular phylogenetic analysis was conducted using two fragments of mitochondrial DNA, cytochrome b (cyt b, 891 bp) and NADH dehydrogenase subunit 2 (ND2, 627 bp) genes. Phylogenetic trees of cyt b revealed that all Korean specimens formed a monophyletic group with Japanese scops owl Otus semitorques with very low sequence divergence (d=0.008). We obtained a similar ND2 tree as well (d=0.003); however, the genetic distance between Korean individuals and O. lempiji from GenBank (AJ004026-7, EU348987, and EU601036) was very high and sufficient enough to separate them as species (cyt b, d=0.118; ND2, d=0.113). We also found that Korean species showed high differentiation from O. bakkamoena (AJ004018-20 and EU601034; cyt b, d=0.106; ND2, d=0.113) and O. lettia (EU601109 and EU601033, cyt b, d=0.110; ND2, d=0.117) as well. Therefore, we suggest that the Korean collared scops owl should be designated as Otus semitorques.

• Molecules and Cells
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• 제22권2호
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• pp.146-153
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• 2006

Cell polarity is critical for the division, differentiation, migration, and signaling of eukaryotic cells. RAX2 of budding yeast encodes a membrane protein localized at the cell cortex that helps maintain the polarity of the bipolar pattern. Here, we designate SPAC6f6.06c as $rax2^+$ of Schizosaccharomyces pombe, based on its sequence homology with RAX2, and examine its function in cell polarity. S. pombe $rax2^+$ is not essential, but ${\Delta}rax2$ cells are slightly smaller and grow slower than wild type cells. During vegetative growth or arrest at G1 by mutation of cdc10, deletion of $rax2^+$ increases the number of cells failing old end growth just after division. In addition, this failure of old end growth is dramatically increased in ${\Delta}tea1{\Delta}rax2$, pointing to genetic interaction of $rax2^+$ with $tea1^+$. ${\Delta}rax2$ cells contain normal actin and microtubule cytoskeletons, but lack actin cables, and the polarity factor for3p is not properly localized at the growing tip. In ${\Delta}rax2$ cells, and endogenous rax2p is localized at the cell cortex of growing cell tips in an actin- and microtubule-dependent manner. However, ${\Delta}rax2$ cells show no defects in cell polarity during shmoo formation and conjugation. Taken together, these observations suggest that rax2p controls the cell polarity of fission yeast during vegetative growth by regulating for3p localization.

• Genomics & Informatics
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• 제13권4호
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• pp.137-145
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• 2015

Selective sweep can cause genetic differentiation across populations, which allows for the identification of possible causative regions/genes underlying important traits. The pig has experienced a long history of allele frequency changes through artificial selection in the domestication process. We obtained an average of 329,482,871 sequence reads for 24 pigs from three pig breeds: Yorkshire (n = 5), Landrace (n = 13), and Duroc (n = 6). An average read depth of 11.7 was obtained using whole-genome resequencing on an Illumina HiSeq2000 platform. In this study, cross-population extended haplotype homozygosity and cross-population composite likelihood ratio tests were implemented to detect genes experiencing positive selection for the genome-wide resequencing data generated from three commercial pig breeds. In our results, 26, 7, and 14 genes from Yorkshire, Landrace, and Duroc, respectively were detected by two kinds of statistical tests. Significant evidence for positive selection was identified on genes ST6GALNAC2 and EPHX1 in Yorkshire, PARK2 in Landrace, and BMP6, SLA-DQA1, and PRKG1 in Duroc. These genes are reportedly relevant to lactation, reproduction, meat quality, and growth traits. To understand how these single nucleotide polymorphisms (SNPs) related positive selection affect protein function, we analyzed the effect of non-synonymous SNPs. Three SNPs (rs324509622, rs80931851, and rs80937718) in the SLA-DQA1 gene were significant in the enrichment tests, indicating strong evidence for positive selection in Duroc. Our analyses identified genes under positive selection for lactation, reproduction, and meat-quality and growth traits in Yorkshire, Landrace, and Duroc, respectively.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제38권2호
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• pp.101-109
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• 2012

Objectives: The inactivation of the tumor suppressor gene $p16^{INK4a}$ plays an important role in the development of malignant tumors, including oral squamous cell carcinoma. The p16 gene is involved in the p16/cyclin-dependent kinase/retinoblastoma (Rb) gene pathway of cell cycle control. The p16 protein is considered a negative regulator of this pathway. The p16 gene encodes an inhibitor of cyclin-dependent kinases 4 and 6 which regulate the phosphorylation of the retinoblastoma gene and G1 to S phase transition in the cell cycle. However, the p16 gene can lose its functionality through point mutations, loss of heterozygosity or methylation of its promoter region. Materials and Methods: In this study, the authors analyzed the correlation between various clinicopathological findings- patient age, gender and smoking, disease recurrence, tumor size, stage, and differentiation- and p16 protein expression or p16 promoter hypermethylation in 59 cases of head and neck squamous cell carcinoma. Results: The results revealed p16 protein expression and p16 promoter hypermethylation in 28 cases (47.5%) and 21 cases (35.6%), respectively, of head and neck squamous cell carcinoma. However, neither p16 protein expression nor p16 promoter hypermethylation had any statistical influence on clinicopathological findings or survival rate. Conclusion: This data, and a review of the literature, suggest that p16 promoter hypermethylation cannot yet be used as an independent prognostic factor influencing carcinogenesis, but must be considered as an important factor along with other genetic alterations affecting the pRb pathway.

• 한방안이비인후피부과학회지
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• 제15권1호
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• pp.226-252
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• 2002

The results of a literature study about the comparison of Oriental-Occidental medicine on the Atopic dennatitis were as follows. 1. In Oriental medicine, Atopic dermatitis belongs to the category of the Naesun(내선), Taeryumchang(胎斂瘡), Eczema(濕疹), Chimumchang(浸淫瘡). In Occidental medicine, the other names of Atopic dermatitis are allergic eczema, IgE dermatitis, flexual eczema etc. 2. In Oriental medicine, the definition of atopic dermatitis includes chang(瘡), sun(선), and pung(風). Occidental medicine, is one of the intrinsic eczema classifications. In fact eczema term circumscribes dermatitis in atopic patients. 3. In Oriental medicine, the etiology and pathogenesis of Atopic dermatitis arose from the state of cogenital defect(稟賦不足), internal accumulation of damp and heat(濕熱內蘊) at first, and then invaded pathogenic wind, damp, heat factors again, and combined endo-exoteric pathogenic factors. So it appears in skin. In Occidental medicine, the etiology and pathogenesis of Atopic dermatitis approaches in genetic, allergic and immunologic, pharmacophysioloic aspects. It is only a hypothesis but there is no known facts about radical aetiology. 4. In Oriental medicine, differentiation of syndromes classifies manifestation aspects, etiology and pathogenesis, and invasion period. In Occidental medicine, it divides into an invasion period, and clinical aspect etc. 5. In Oriental medicine, Internal theraphys of Atopic dermatitis are decoction of ingredients(湯劑), pills(丸), and tablet(片). So, it prescribes as treatments on the ground of differentition of syndrome. In Occidental medicine, there is no radical therapy because Atopic dispositions don't change. But steroid, antihistamine as symptomatic tre atments are generally used in Occidental medicine. 6. In Oriental medicine, external therapies are wet dressings(濕敷), lotion(洗劑), powder(散劑), adhesive plasters(膏劑), oil(油劑), smoking(烟薰法), warm over fire therapy(熱烘療法), acupunture and moxibustion therapy(鍼灸療法).

• 한국수자원학회논문집
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• 제46권4호
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• pp.389-400
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• 2013

이 논문에서는 관망시스템의 마찰항을 보정하기 위해서, 부정류 마찰 모형과 Levenberg Marquardt 방법을 합성하였다. 부정류 마찰항을 고려하기 위한 방법으로 빈도 의존 마찰항을 사용하였으며, 특성선 방법을 모형 개발의 기반으로 하였다. 최적화에 필요한 Hessian과 Jacobian 행렬을 구하기위해서 수압을 직접 마찰항에 미분한 항을 계산하였으며, 특성선 방법상에서의 다양한 수압과 유량에 대한 마찰계수의 민감도를 수식으로 유도하였다. 간단한 관망을 가정한 뒤, 갑작스런 밸브의 거동으로 도입된수압의 시계열을 확보하였고, 이를 이용하여 정상류 마찰 모형과 부정류 마찰모형의 마찰항 보정을 수행하였다. 제안된 방법과 진화 연산 알고리즘의 마찰항 수렴거동을 비교하였으며, Leveberg Marquardt 방법의 안정적이고 신속한 수렴결과를 확인하였다.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1987년도 식물생명공학 심포지움 논문집 Proceedings of Symposia on Plant Biotechnology
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• pp.149-155
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• 1987

Growth and development of a higher plant, or any living organism for that matter, could be defined as an orderly expression of the genome in time and space in close interaction with the environment. During differentiation and development of a tissue or organ a group of genes must be selectively turned on or turned off mainly by trans-acting regulators. In this general concept of regulation of regulation of gene expression, a DNA molecule is recognized at a specific nucleotide sequence by DNA-binding factors. Molecular biology of the regulatory factors such as hormones, and their receptors, target DNA sequences and DNA-binding proteins are well advanced. What is not clearly understood is the molecular basis of the interactions between DNA and binding factors, expecially of the usages of the dyad symmetry of the target DNA sequences and the dimeric nature of the DNA-binding proteins. A unique 3-dimensional structure of DNA has been proposed that may play an important role in the orderly expression of the gene. A foldback intercoil (FBI) DNA configuration which was originally found by electron microscopy among mtDNA molecules from pearl millet has some unique features. The FBI configuration of DNA is believed to be formed when a flexible double helix folds back and interwines in the widened major grooves resulting in a four stranded, intercoil DNA whose thickness is the same as that of double stranded DNA. More recently, the FBI structure of DNA has been also induced in vitro by a novel enzyme which was purified from pearl millet mitochondria. It has been proposed that the FBI DNA could be utillized in intramolecular recombination which leads to inversion or deletion, and in intermolecular recombination which can lead to either site-specific recombination, genetic recombination via single strand invasion, or cross strand recombination. The structure and function of DNA in 3-dimensional aspect is emphasized for better understanding orderly expression of genes during growth and development.