• Asian-Australasian Journal of Animal Sciences
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• v.31 no.4
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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.

• The Plant Pathology Journal
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• v.26 no.1
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• pp.70-79
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• 2010

Rice blast is one of the serious devastating diseases. This study was carried out to determine the genetic diversities of blast resistance (R) genes form 86 accessions of aromatic rice with eight SNP markers, z4792, zt4792, z60510, zt6057, k6415, k6411, k39575 and t256, which showed the close-set linkage to 6 major genes, Piz, Piz-t, Pik, Pik-m, Pik-p, and Pit. Four accessions of indica type, Mayataung, Yekywin Yinkya Hmwe, Basmati9-93, and Basmati5854, showed the positive amplicons of six major genes. Among 86 accessions, 83 accessions were detected both or one of Piz and Piz-t genes. Seventy three accessions contained the Piz gene with z4792 marker. In addition, 30 and 71 accessions possessed Piz-t gene with zt4792 and zt6057 markers, respectively. Ten accessions showed the positive bands for the Piz-t gene with both zt4792 and zt6057 markers. Only one accession, Khau Nua Keo, was not amplified for both Piz and Piz-t gene. But japonica type, Gerdeh, possessed only Piz gene between Piz and Piz-t. Fifty two accessions showed the three of Pik multiple genes and Pit gene. Four accessions, Iari7447, Daebunhyangdo2, Shiyayuuine, and Basmati 6129 possessed a Pik-p gene. Especially, Pit gene on chromosome 1 was detected with t256 marker in all of 83 accessions, exception of A-2, one accession of japonica type.

• Journal of the Korean Data and Information Science Society
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• v.24 no.6
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• pp.1331-1339
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• 2013

The lipoprotein lipase (LPL) gene can be considered a functional candidate gene that regulates fatty acid composition. Oh etc (2013) investigated the relationship between unsaturated fatty acids and five novel SNPs, and had confirmed that three polymorphic SNPs (c.322G>A, c.329A>T and c.1591G>A) were associated with fatty acid composition. We have used generalized linear model for adjusted environmental effects and multifactor dimensionality reduction (MDR) method to identify gene-gene interaction effect of statistical model in general. We applied the MDR method on the identify major interaction effects of exonic single nucleotide polymorphisms (SNPs) in the LPL gene for economic traits in Korean cattles population.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.8
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• pp.1088-1097
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• 2005

Main objectives of this study were to investigate accuracy, bias and power of linear and threshold model segregation analysis methods for detection of major genes in categorical traits in farm animals. Maximum Likelihood Linear Model (MLLM), Bayesian Linear Model (BALM) and Bayesian Threshold Model (BATM) were applied to simulated data on normal, categorical and binary scales as well as to disease data in pigs. Simulated data on the underlying normally distributed liability (NDL) were used to create categorical and binary data. MLLM method was applied to data on all scales (Normal, categorical and binary) and BATM method was developed and applied only to binary data. The MLLM analyses underestimated parameters for binary as well as categorical traits compared to normal traits; with the bias being very severe for binary traits. The accuracy of major gene and polygene parameter estimates was also very low for binary data compared with those for categorical data; the later gave results similar to normal data. When disease incidence (on binary scale) is close to 50%, segregation analysis has more accuracy and lesser bias, compared to diseases with rare incidences. NDL data were always better than categorical data. Under the MLLM method, the test statistics for categorical and binary data were consistently unusually very high (while the opposite is expected due to loss of information in categorical data), indicating high false discovery rates of major genes if linear models are applied to categorical traits. With Bayesian segregation analysis, 95% highest probability density regions of major gene variances were checked if they included the value of zero (boundary parameter); by nature of this difference between likelihood and Bayesian approaches, the Bayesian methods are likely to be more reliable for categorical data. The BATM segregation analysis of binary data also showed a significant advantage over MLLM in terms of higher accuracy. Based on the results, threshold models are recommended when the trait distributions are discontinuous. Further, segregation analysis could be used in an initial scan of the data for evidence of major genes before embarking on molecular genome mapping.

• Journal of fish pathology
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• v.32 no.2
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• pp.49-57
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• 2019

In this study, we collected 39 megalocytiviruses isolated from diseased fish in Korea from 2012 to 2018. Major capsid protein (MCP) gene, a part of vascular endothelial growth factor (VEGF) gene and histidine triad motif-like protein (HIT) genes of Megalocytivirus were targeted for PCR amplification and analysis of those DNA nucleotide sequences. Korean strains revealed two genotypes (red sea bream iridovirus and turbot reddish body iridovirus types) based on the phylogeny of MCP gene. The red sea bream iridovirus type (RSIV-type) megalocytiviruses were divided into RSIV-subgroup 1 and 2. From the phylogenetic analysis of the VEGF genes, a genotypic variant of RSIV-type Megalocytivirus was identified. The HIT-like protein gene was detected in RSIVs, but not in TBRIV and ISKNV, suggesting that HIT-like protein gene may be specific in RSIV.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.6
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• pp.871-884
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• 2009

Gene expression profiling is a useful tool for identifying critical genes and pathways in metabolism. The objective of this study was to determine the major differences in the expression of genes associated with metabolism and metabolic regulation in liver and mammary tissues of lactating cows. We used the Michigan State University bovine metabolism (BMET) microarray; previously, we have designed a bovine metabolism-focused microarray containing known genes of metabolic interest using publicly available genomic internet database resources. This is a high-density array of 70mer oligonucleotides representing 2,349 bovine genes. The expression of 922 genes was different at p<0.05, and 398 genes (17%) were differentially expressed by two-fold or more with 222 higher in liver and 176 higher in mammary tissue. Gene ontology categories with a high percentage of genes more highly expressed in liver than mammary tissues included carbohydrate metabolism (glycolysis, glucoenogenesis, propanoate metabolism, butanoate metabolism, electron carrier and donor activity), lipid metabolism (fatty acid oxidation, chylomicron/lipid transport, bile acid metabolism, cholesterol metabolism, steroid metabolism, ketone body formation), and amino acid/nitrogen metabolism (amino acid biosynthetic process, amino acid catabolic process, urea cycle, and glutathione metabolic process). Categories with more genes highly expressed in mammary than liver tissue included amino acid and sugar transporters and MAPK, Wnt, and JAK-STAT signaling pathways. Real-time PCR analysis showed consistent results with those of microarray analysis for all 12 genes tested. In conclusion, microarray analyses clearly identified differential gene expression profiles between hepatic and mammary tissues that are consistent with the differences in metabolism of these two tissues. This study enables understanding of the molecular basis of metabolic adaptation of the liver and mammary gland during lactation in bovine species.

• Macromolecular Research
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• v.15 no.2
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• pp.100-108
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• 2007

Gene therapy has become a promising strategy for the treatment of genetically based diseases, such as cancer, which are currently considered incurable. A major obstacle in the field of cancer gene therapy is the development of a safe and efficient delivery system for therapeutic gene transfer. Non-viral vectors have attracted great interest, as they are simple to prepare, stable, easy to modify and relatively safe compared to viral vectors. In this review, an insight into the strategies developed for polyethylenimine (PEI)-based non-viral vectors has been provide, including improvement of the polyplex properties by incorporating hydrophilic spacer, poly(ethylene glycol) (PEG). Moreover, this review will summarize the strategies for the tumor targeting. Specifically, a targeted polymeric gene delivery system, PEI-g-PEG-RGD, will be introduced as an efficient gene delivery vector for tumor therapy, including its functional analysis both in vitro and in vivo.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.7
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• pp.938-943
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• 2016

Development of body hair is an important physiological and cellular process that leads to better adaption in tropical environments for dairy cattle. Various studies suggested a major gene and, more recently, associated genes for hairy locus in dairy cattle. Main aim of this study was to i) employ a variant of the discordant sib pair model, in which half sibs from the same sires are randomly sampled using their affection statues, ii) use various single marker regression approaches, and iii) use whole genome regression approaches to dissect genetic architecture of the hairy gene in the cattle. Whole and single genome regression approaches detected strong genomic signals from Chromosome 23. Although there is a major gene effect on hairy phenotype sourced from chromosome 23: whole genome regression approach also suggested polygenic component related with other parts of the genome. Such a result could not be obtained by any of the single marker approaches.

• Parasites, Hosts and Diseases
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• v.55 no.4
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• pp.367-374
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• 2017

Despite the broad distribution of leishmaniasis among Iranians and animals across the country, little is known about the genetic characteristics of the causative agents. Applying both HSP70 PCR-RFLP and sequence analyses, this study aimed to evaluate the genetic diversity and phylogenetic relationships among Leishmania spp. isolated from Iranian endemic foci and available reference strains. A total of 36 Leishmania isolates from almost all districts across the country were genetically analyzed for the HSP70 gene using both PCR-RFLP and sequence analysis. The original HSP70 gene sequences were aligned along with homologous Leishmania sequences retrieved from NCBI, and subjected to the phylogenetic analysis. Basic parameters of genetic diversity were also estimated. The HSP70 PCR-RFLP presented 3 different electrophoretic patterns, with no further intraspecific variation, corresponding to 3 Leishmania species available in the country, L. tropica, L. major, and L. infantum. Phylogenetic analyses presented 5 major clades, corresponding to 5 species complexes. Iranian lineages, including L. major, L. tropica, and L. infantum, were distributed among 3 complexes L. major, L. tropica, and L. donovani. However, within the L. major and L. donovani species complexes, the HSP70 phylogeny was not able to distinguish clearly between the L. major and L. turanica isolates, and between the L. infantum, L. donovani, and L. chagasi isolates, respectively. Our results indicated that both HSP70 PCR-RFLP and sequence analyses are medically applicable tools for identification of Leishmania species in Iranian patients. However, the reduced genetic diversity of the target gene makes it inevitable that its phylogeny only resolves the major groups, namely, the species complexes.

• The Plant Pathology Journal
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• v.31 no.1
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• pp.12-24
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• 2015

Rice Blast is the most devastating disease causing major yield losses in every year worldwide. It had been proved that using resistant rice varieties would be the most effective way to control this disease. Molecular screening and genetic diversities of major rice blast resistance genes were determined in 192 rice germplasm accessions using simple sequence repeat (SSR) markers. The genetic frequencies of the 10 major rice blast resistance genes varied from 19.79% to 54.69%. Seven accessions IC337593, IC346002, IC346004, IC346813, IC356117, IC356422 and IC383441 had maximum eight blast resistance gene, while FR13B, Hourakani, Kala Rata 1-24, Lemont, Brown Gora, IR87756-20-2-2-3, IC282418, IC356419, PKSLGR-1 and PKSLGR-39 had seven blast resistance genes. Twenty accessions possessed six genes, 36 accessions had five genes, 41 accessions had four genes, 38 accessions had three genes, 26 accessions had two genes, 13 accessions had single R gene and only one accession IC438644 does not possess any one blast resistant gene. Out of 192 accessions only 17 accessions harboured 7 to 8 blast resistance genes.