• Title/Summary/Keyword: Nucleotide sequencing

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Genetic diversity of spotted scat (Scatophagus argus) in Vietnam based on COI genes

  • Huy Van Nguyen;Minh Tu Nguyen;Nghia Duc Vo;Nguyen Thi Thao Phan;Quang Tan Hoang
    • Fisheries and Aquatic Sciences
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    • v.25 no.12
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    • pp.637-647
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    • 2022
  • A spotted scat, Scatophagus argus, has a high nutritional value and is among Asia's most widely consumed fish species. Thua Thien Hue's consumption market considers this species to be of high economic value and requires protection and conservation of the population. However, the studies on the identification and genetic diversity of S. argus distributed in Vietnam are still lacking. Therefore, mitochondrial cytochrome c oxidase subunit I (COI) gene was utilized to distinguish different populations and investigate the genetic diversity of two populations of S. argus from Tam Giang lagoon, Thua Thien Hue province (n = 31) and Ca Mau province (n = 14). The sequencing results indicated 13 distinct haplotypes among 45 sequences. Five single nucleotide polymorphisms were observed to distinguish Hue spotted scat population. The S. argus population in Ca Mau province was higher haplotype diversity (Hd) and nucleotide diversity (π) than those of Thua Thien Hue province, which demonstrates that there are minor differences between haplotypes. There were genetic distances ranging from 0%-4% within the populations and 6.67% between the two populations. In addition to the sequencing, the comparison of morphology, biology, culture, and the growth rate was sufficient to distinguish the spotted scat S. argus in Thua Thien Hue from Ca Mau.

Association between Single Nucleotide Polymorphisms of the Major Histocompatibility Complex Class II Gene and Newcastle Disease Virus Titre and Body Weight in Leung Hang Khao Chickens

  • Molee, A.;Kongroi, K.;Kuadsantia, P.;Poompramun, C.;Likitdecharote, B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.1
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    • pp.29-35
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    • 2016
  • The aim of the present study was to investigate the effect of single nucleotide polymorphisms in the major histocompatibility complex (MHC) class II gene on resistance to Newcastle disease virus and body weight of the Thai indigenous chicken, Leung Hang Khao (Gallus gallus domesticus). Blood samples were collected for single nucleotide polymorphism analysis from 485 chickens. Polymerase chain reaction sequencing was used to classify single nucleotide polymorphisms of class II MHC. Body weights were measured at the ages of 3, 4, 5, and 7 months. Titres of Newcastle disease virus at 2 weeks to 7 months were determined and the correlation between body weight and titre was analysed. The association between single nucleotide polymorphisms and body weight and titre were analysed by a generalized linear model. Seven single nucleotide polymorphisms were identified: C125T, A126T, C209G, C242T, A243T, C244T, and A254T. Significant correlations between log titre and body weight were found at 2 and 4 weeks. Associations between single nucleotide polymorphisms and titre were found for C209G and A254T, and between all single nucleotide polymorphisms (except A243T) and body weight. The results showed that class II MHC is associated with both titre of Newcastle disease virus and body weight in Leung Hang Khao chickens. This is of concern because improved growth traits are the main goal of breeding selection. Moreover, the results suggested that MHC has a pleiotropic effect on the titre and growth performance. This mechanism should be investigated in a future study.

Screening of the Single Nucleotide Polymorphisms in the Protamine 1 and 2 Genes of Korean Infertile Men (한국 남성 불임환자에서 Protamine 1과 2 유전자의 Single Nucleotide Polymorphism에 관한 연구)

  • Lee, Hyoung-Song;Choi, Hye Won;Park, Yong-Seog;Seo, Ju Tae;Koong, Mi Kyoung;Jun, Jin Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.3
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    • pp.279-286
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    • 2005
  • Objective: Although several genetic factors have been associated with defects in human spermatogenesis, the unambiguous causative genes have not been elucidated. The male infertility by haploinsufficiency of PRM1 or PRM2 has been reported in mouse model. The aim of this study was to identify the single nucleotide polymorphisms (SNPs) of PRM1 and PRM2, related to the genotype of Korean infertile men. Methods: Genomic DNAs were extracted from peripheral bloods of infertile men with oligozoospermia or azoospermia, and analyzed using polymerase chain reaction (PCR) and direct sequencing. We carried out the direct sequencing analysis of amplified fragments in PRM1 (557 nucleotides from -42 to 515) and PRM2 (599 nucleotides from 49 to 648) genes, respectively. Results: Three SNPs of coding region in the PRM1 gene was found in the analysis of 130 infertile men. However, the SNPs at a133g (aa 96.9%, ag 3.1% and gg 0.0%), c160a (cc 99.2%, ca 0.8% and aa 0.0%) and c321a (cc 56.9%, ca 35.4% and aa 7.7%) coded the same amino acids, in terms of silence phenotypes. On the other hand, as results of the PRM2 gene sequencing in 164 infertile men, only two SNPs, g398c (gg 62.2%, gc 31.1% and ga 6.7%) and a473c (aa 63.4%, ac 29.9% and cc 6.7%), were identified in the intron of the PRM2 gene. Conclusions: There was no mutation and significant SNPs on PRM1 and PRM2 gene in Korean infertile men. These results suggest that the PRM1 and PRM2 genes are highly conserved and essential for normal fertility of men.

A case of interdigitating dendritic cell sarcoma studied by whole-exome sequencing

  • Hong, Ki Hwan;Song, Soyoung;Shin, Wonseok;Kang, Keunsoo;Cho, Chun?Sung;Hong, Yong Tae;Han, Kyudong;Moon, Jeong Hwan
    • Genes and Genomics
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    • v.40 no.12
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    • pp.1279-1285
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    • 2018
  • Interdigitating dendritic cell sarcoma (IDCS) is an aggressive neoplasm and is an extremely rare disease, with a challenging diagnosis. Etiology of IDCS is also unknown and most studies with only case reports. In our case, immunohistochemistry showed that the tumor cells were positive for S100, CD45, and CD68, but negative for CD1a and CD21. This study aimed to investigate the causative factors of IDCS by sequencing the protein-coding regions of IDCS. We performed whole-exome sequencing with genomic DNA from blood and sarcoma tissue of the IDCS patient using the Illumina Hiseq 2500 platform. After that, we conducted Sanger sequencing for validation of sarcoma-specific variants and gene ontology analysis using DAVID bioinformatics resources. Through comparing sequencing data of sarcoma with normal blood, we obtained 15 nonsynonymous single nucleotide polymorphisms (SNPs) as sarcoma-specific variants. Although the 15 SNPs were not validated by Sanger sequencing due to tumor heterogeneity and low sensitivity of Sanger sequencing, we examined the function of the genes in which each SNP is located. Based on previous studies and gene ontology database, we found that POLQ encoding DNA polymerase theta enzyme and FNIP1 encoding tumor suppressor folliculin-interacting protein might have contributed to the IDCS. Our study provides potential causative genetic factors of IDCS and plays a role in advancing the understanding of IDCS pathogenesis.

PCR-SSCP Polymorphism of Inhibin ${\beta}_A$ Gene in Some Sheep Breeds

  • Chu, M.X.;Xiao, C.T.;Fu, Y.;Fang, L.;Ye, S.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.7
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    • pp.1023-1029
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    • 2007
  • Inhibins participate in the regulation of pituitary follicle-stimulating hormone synthesis and secretion, follicular maturation and steroidogenesis in the female. Inhibin ${\beta}_A$ gene (INHBA) was studied as a candidate gene for the prolificacy of sheep. Single nucleotide polymorphisms of the entire coding region and partial 3' untranslated region of INHBA were detected by PCR-SSCP in two high fecundity breeds (Small Tail Han and Hu sheep) and six low fecundity breeds (Dorset, Texel, German Mutton Merino, South African Mutton Merino, Chinese Merino and Corriedale sheep). Only the PCR products amplified by primers 3, 4 and 5 displayed polymorphisms. For primer 3, genotype CC was only detected in Chinese Merino sheep, genotype AA was detected in the other seven sheep breeds. Genotype BB was only detected in Hu sheep. Only Hu sheep displayed polymorphism. Eight or four nucleotide mutations were revealed between BB or CC and AA, respectively, and these mutations did not result in any amino acid change. For primer 4, genotypes EE, EG and GG were detected in Dorset and German Mutton Merino sheep, genotypes EE, EF and FF were detected in Chinese Merino sheep, only genotype EE was detected in the other five sheep breeds. Only Dorset, German Mutton Merino and Chinese Merino sheep displayed polymorphism. Sequencing revealed one nucleotide mutation ($114G{\rightarrow}A$) of exon 2 of INHBA gene between genotype FF and genotype EE, and this mutation did not cause any amino acid change. Another nucleotide change ($143C{\rightarrow}T$) was identified between genotype GG and genotype EE, and this mutation resulted in an amino acid change of $serine{\rightarrow}leucine$. For primer 5, genotypes KK and KL were detected in German Mutton Merino and Corriedale sheep, genotypes KK, LL and KL were detected in the other six sheep breeds. Genotype MM was only detected in Hu sheep. All of these eight sheep breeds displayed polymorphism. Sequencing revealed one nucleotide mutation ($218A{\rightarrow}G$) of exon 2 of the INHBA gene between genotype LL and genotype KK, and nine nucleotide mutations between genotype MM and genotype KK. These mutations did not alter amino acid sequence. The partial sequence (395 bp for exon 1 and 933 bp for exon 2) of the INHBA gene in Small Tail Han sheep (with genotype KK for primer 5) was submitted into GenBank (accession number EF192431). Small Tail Han sheep displayed polymorphisms only in the fragment amplified by primer 5. The Small Tail Han ewes with genotype LL had 0.53 (p<0.05) or 0.63 (p<0.05) more lambs than those with genotype KL or KK, respectively. The Small Tail Han ewes with genotype KL had 0.10 (p>0.05) more lambs than those with genotype KK.

Shallow Population Genetic Structures of Thread-sail Filefish (Stephanolepis cirrhifer) Populations from Korean Coastal Waters

  • Yoon, M.;Park, W.;Nam, Y.K.;Kim, D.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.2
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    • pp.170-176
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    • 2012
  • Genetic diversities, population genetic structures and demographic histories of the thread-sail filefish Stephanolepis cirrhifer were investigated by nucleotide sequencing of 336 base pairs of the mitochondrial DNA (mtDNA) control region in 111 individuals collected from six populations in Korean coastal waters. A total of 70 haplotypes were defined by 58 variable nucleotide sites. The neighbor-joining tree of the 70 haplotypes was shallow and did not provide evidence of geographical associations. Expansion of S. cirrhifer populations began approximate 51,000 to 102,000 years before present, correlating with the period of sea level rise since the late Pleistocene glacial maximum. High levels of haplotype diversities ($0.974{\pm}0.029$ to $1.000{\pm}0.076$) and nucleotide diversities (0.014 to 0.019), and low levels of genetic differentiation among populations inferred from pairwise population FST values (-0.007 to 0.107), support an expansion of the S. cirrhifer population. Hierarchical analysis of molecular variance (AMOVA) revealed weak but significant genetic structures among three groups ($F_{CT}$ = 0.028, p<0.05), and no genetic variation within groups (0.53%; $F_{SC}$ = 0.005, p = 0.23). These results may help establish appropriate fishery management strategies for stocks of S. cirrhifer and related species.

Sequence of the spike gene containing antigenic sites A and D of transmissible gastroenteritis virus isolated in Korea (국내분리 돼지 전염성 위장염 바이러스의 antigenic site A와 D를 포함하는 spike gene의 염기서열 분석)

  • Kwon, Hyuk-moo;Pi, Jae-ho;Seong, Hwan-woo
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.319-327
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    • 1998
  • The nucleotide sequences of spike (S) glycoprotein containing antigenic sites A and D of TGEV isolated in Korea were determined and compared with published sequences for TGEVs. The TGEV 133 and DAE5 strains had 97.40% nucleotide sequence similarity. The overall nucleotide sequence similarity of the 133 and DAE5 strains compared with other TGEV strains was between 96.86% and 99.15%. The similarity of the predicted amino acid sequence of the 133 and DAE5 strains was 94.93%. The TGEV 133 and DAE5 strains had 94.93-98.61% amino acid similarity with published sequences of other TGEV strains. The sequences of amino acid codons in the antigenic sites A and D were identical among all the viruses although there were several nucleotide changes in region containing antigenic sites A and D of Korean TGEV isolates. By phylogenetic analysis of the sequences, two Korean isolates 133 and DAE5 seemed to be derived from different lineages. These studies showed that a distinct difference in genome exists among TGEV field strains isolated in Korea.

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Association between Single Nucleotide Polymorphisms of Fatty Acid Synthase and Fat Deposition in the Liver of the Overfed Goose

  • Wu, Wei;Guo, Xuan;Zhang, Lei;Hu, Dan
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.9
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    • pp.1244-1249
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    • 2014
  • Goose fatty liver is one of the most delicious and popular foods in the world, but there is no reliable genetic marker for the early selection and breeding of geese with good liver-producing potential. In our study, one hundred and twenty-four 78-day-old Landes geese bred in Shunda Landes goose breeding farm, Jiutai, Jilin, China were selected randomly. The fatty livers were sampled each week after overfeeding during a three week period. Polymerase chain reaction-single strand conformation polymorphism and DNA sequencing were used to identify single nucleotide polymorphisms (SNPs) of fatty acid synthase (FAS), which is an important enzyme involved in the synthesis of fat under both physiological and pathological conditions. Least-squares correlation was established between these SNPs and fatty liver weight, abdominal fat weight, and intestinal fat weight of the overfed Landes geese, respectively. The results showed that fatty liver weight of geese with EF and FF genotypes (amplified by primer P1) was significantly higher than that of the EE genotype (p<0.05), and liver weight of CD and DD genotypes (amplified by primer P2) was significantly higher than that of the CC genotype (p<0.05). Different genotype combinations showed different liver weights, and from highest to lowest were ABDD, DDEF, DDFF, DDEE, ABEF, ABFF, AADD, and CDEF. Further analysis of DNA sequencing showed that there were two SNPs within the 5' promoter region the FAS gene. The geese of EF and FF genotypes carried a change of T to C, and the geese of CD and DD genotypes carried a change of A to G. The changes of the bases could potentially influence the binding of some transcription factors to this region as to regulate FAS gene. To our knowledge, this is the first report of SNPs found within the 5' promoter region of the Landes goose FAS gene, and our data will provide an insight for early selection of geese for liver production.

Transcriptome analysis, microsatellite marker information, and orthologous analysis of Capsicum annuum varieties

  • Ahn, Yul-Kyun;Karna, Sandeep;Kim, Jeong-Ho;Lee, Hye-Eun;Kim, Jin-Hee;Kim, Do-Sun
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.311-316
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    • 2016
  • The efficacy of plant breeding has been enhanced by application of molecular markers in population screening and selection. Pepper (Capsicum annuum L.) is a major staple crop that is economically important with worldwide distribution. It is valued for its spicy taste and medicinal effect. The aim of this study was to discover single nucleotide polymorphisms (SNPs), microsatellite markers information, and percentage sharing through orthologous analysis of pepper-specific pungency-related genes. Here, we report the results of transcriptome analysis and microsatellite markers for four pepper varieties that possess a pungency-related gene. Orthologous analyses was performed to identify species-specific pungency-related genes in pepper, Arabidopsis thaliana L., potato (Solanum tuberosum L.), and tomato (Solanum lycopersicum L.). Advancements in next-generation sequencing technologies enabled us to quickly and cost-effectively assemble and characterize genes to select molecular markers in various organisms, including pepper. We identified a total of 9762, 7302, 8596, and 6886 SNPs for the four pepper cultivars Blackcluster, Mandarine, Saengryeg 211, and Saengryeg 213, respectively. We used 454 GS-FLX pyrosequencing to identify microsatellite markers and tri-nucleotide repeats (54.4%), the most common repeats, followed by di-, hexa-, tetra-, and penta-nucleotide repeats. A total of 5156 (15.9%) pepper-specific pungency-related genes were discovered as a result of orthologous analysis.

In silico approaches to discover the functional impact of non-synonymous single nucleotide polymorphisms in selective sweep regions of the Landrace genome

  • Shin, Donghyun;Won, Kyung-Hye;Song, Ki-Duk
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.12
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    • pp.1980-1990
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    • 2018
  • Objective: The aim of this study was to discover the functional impact of non-synonymous single nucleotide polymorphisms (nsSNPs) that were found in selective sweep regions of the Landrace genome Methods: Whole-genome re-sequencing data were obtained from 40 pigs, including 14 Landrace, 16 Yorkshire, and 10 wild boars, which were generated with the Illumina HiSeq 2000 platform. The nsSNPs in the selective sweep regions of the Landrace genome were identified, and the impacts of these variations on protein function were predicted to reveal their potential association with traits of the Landrace breed, such as reproductive capacity. Results: Total of 53,998 nsSNPs in the mapped regions of pigs were identified, and among them, 345 nsSNPs were found in the selective sweep regions of the Landrace genome which were reported previously. The genes featuring these nsSNPs fell into various functional categories, such as reproductive capacity or growth and development during the perinatal period. The impacts of amino acid sequence changes by nsSNPs on protein function were predicted using two in silico SNP prediction algorithms, i.e., sorting intolerant from tolerant and polymorphism phenotyping v2, to reveal their potential roles in biological processes that might be associated with the reproductive capacity of the Landrace breed. Conclusion: The findings elucidated the domestication history of the Landrace breed and illustrated how Landrace domestication led to patterns of genetic variation related to superior reproductive capacity. Our novel findings will help understand the process of Landrace domestication at the genome level and provide SNPs that are informative for breeding.