• Title/Summary/Keyword: RFLP.

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Genetic Variation of Growth Hormone Gene and Its Relationship with Milk Production Traits in China Holstein Cows

  • Zhou, Guo-li;Zhu, Qi;Jin, Hai-guo;Guo, Shan-li
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.315-318
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    • 2006
  • Associations were analysed between polymorphisms localized in intron 3 of the growth hormone gene (GH-MspI) and milk production traits of 543 China Holstein cows. A PCR-RFLP method was used for identification of genotypes. The following frequencies of genotypes and alleles were found: 0.77, 0.21 and 0.02 for +/+, +/- and -/-, respectively, and 0.87 and 0.13 for $GH^+$ and $GH^-$, respectively. Significant differences between herds were observed in the frequencies of both genotypes and alleles. The results of least squares analysis showed that in all three lactation phases the GH +/+ cows yielded most milk (p<0.01 for lactation I and p<0.05 for lactations II and III), whereas +/- cows showed higher milk fat content than +/+ individuals (p<0.05 for lactation I and II, and p<0.01 for lactation III). The +/+ cows yielded more fat than +/- individuals (p<0.01 only in lactation I). The +/+ cows yielded more milk protein than +/- individuals (p<0.01 for lactation I, II, and III). The +/+ cows produced milk of higher protein content than that of +/- individuals (p<0.05 only in lactation II). Based on these results, we conclude that the +/+ of GH locus should be the favored genotype in China Holstein cow breeds for use in marker-assisted selection programmes.

Identification and Characterization of Osmotolerant Yeast Isolated from Soy Paste (된장에서 분리된 내염성 효모의 동정 및 특성조사)

  • Byun, Myung-Ok;Lee, Seung-Bum;Koo, Bon-Sung;Song, Jae-Kyeong;Ryu, Jin-Chang;Lee, Du-Hyung
    • The Korean Journal of Mycology
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    • v.27 no.3 s.90
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    • pp.181-186
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    • 1999
  • Osmotolerant yeast isolated from soy paste could grow on media with 2 M NaCl. This strain was identified as Zygosaccharomyces rouxii by biological characteristics, RFLP of ribosomal DNA and mating with compatible haploid strain. Growing rate of the Z. rouxii YDJ was slower than Saccharomyces cerevisiae. Z. rouxii YDJ accumulated trehalose, which is known as one of the osmolytic protectants, in cells cultured on media with salt. Enzyme activity of trehalose phosphate synthase related to trehalose biosynthesis of the YDJ was lower than those of S. cerevisiae. Trehalase activity related trehalose degradation was also lower in Z. rouxii YDJ than S. cerevisiae. However, as Z. rouxii accumulated trehalose by salt treatment, salt tolerancy of Z. rouxii was assumed to be related to trehalose in additon to glycerol.

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Genetic identification of anisakid nematodes isolated from largehead hairtail (Trichiurus japonicus) in Korea

  • Kim, Jeong-Ho;Nam, Woo-Hwa;Jeon, Chan-Hyeok
    • Fisheries and Aquatic Sciences
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    • v.19 no.5
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    • pp.26.1-26.8
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    • 2016
  • Background: The nematode species belonging to genus Anisakis occur at their third larval stage in numerous marine teleost fish species worldwide and known to cause accidental human infection through the ingestion of raw or undercooked fish or squids. They may also draw the attention of consumers because of the visual impact of both alive and dead worms. Therefore, the information on their geographical distribution and clear species identification is important for epidemiological survey and further prevention of human infection. Results: For identification of anisakid nematodes species isolated from largehead hairtail (Trichiurus japonicus), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of internal transcribed spacers of ribosomal DNA were conducted. Mitochondrial cytochrome c oxidase subunit 2 gene was also sequenced, and phylogenetic analysis was conducted. From the largehead hairtail (n = 9), 1259 nematodes were isolated in total. Most of the nematodes were found encapsulated throughout the viscera (56.2 %, 708/1259) or moving freely in the body cavity (41.5 %, 523/1259), and only 0.3 % (4/1259) was found in the muscles. By PCR-RFLP, three different nematode species were identified. Anisakis pegreffii was the most dominantly found (98.7 %, 1243/1259) from the largehead hairtail, occupying 98.7 % (699/708) of the nematodes in the mesenteries and 98.1 % (513/523) in the body cavity. Hybrid genotype (Anisakis simplex ${\times}$ A. pegreffii) occupied 0.5 %, and Hysterothylacium sp. occupied 0.2 % of the nematodes isolated in this study. Conclusions: The largehead hairtail may not significantly contribute accidental human infection of anisakid nematode third stage larvae because most of the nematodes were found from the viscera or body cavity, which are not consumed raw. But, a high prevalence of anisakid nematode larvae in the largehead hairtail is still in concern because they may raise food safety problems to consumers. Immediate evisceration or freezing of fish after catch will be necessary before consumption.

Association between PCR-RFLP Polymorphism of the Fifth Intron in Lipoprotein Lipase Gene and Productive Traits in Pig Resource Family

  • Zhang, B.Z.;Lei, M.G.;Deng, C.Y.;Xiong, Y.H.;Zuo, B.;Li, F.E.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.4
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    • pp.458-462
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    • 2005
  • The study was aimed at detecting polymorphism of the fifth intron in lipoprotein lipase (LPL) gene and analyzing association between the polymorphism and productive traits. A pair of primers was designed for amplifying the fifth intron. Sequence analysis indicated that a G1171C substitution existed in Large White breed. The mutation was detected by PCR-AfaI-RFLP. Polymorphism analysis in a pig resource family showed that there existed significant effects on carcass and meat quality traits. Thoraxwaist fat thickness of BB genotype was significantly higher (14.2%, p<0.05) than that of AA on carcass traits, while BB genotype was significantly lower (3.6% p<0.01, 4.1% p<0.01; 2.3% p<0.01, 1.9% p<0.01; 1.8% p<0.01, 1.4% p<0.05) than AA and AB genotype in pH of m. Longissimus Dorsi (LD), m. Biceps Femoris (BF), m. Semipinali Capitis (SC). The allelic frequencies were also significantly different between indigenous Chinese breeds and exotic breeds. Data analyzed revealed that the mutation locus affected production traits mostly by additive effects. Based on these results, it is necessary to do more studies on LPL gene before making the LPL locus into the application of marker-assisted selection (MAS) programs.

Isolation and Characterization of Strain of Bacillus thuringiensis subsp. kenyae Containing Two Novel cry1-Type Toxin Genes

  • Choi, Jae-Young;Li, Ming Shun;Shim, Hee-Jin;Roh, Jong-Yul;Woo, Soo-Song;Jin, Byung-Rae;Boo, Kyung-Saeng;Je, Yeon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1498-1503
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    • 2007
  • To identify novel crystal proteins, Bacillus thuringiensis 2385-1 was isolated from Korean soil samples and characterized. The H-serotype of 2385-1 was identical to that of subsp. kenyae (H4a4c), and its crystal toxin was bipyramidal-shaped. However, 2385-1 showed a much higher toxicity towards Plutella xylostella and Spodoptera exigua larvae than subsp. kenyae. In addition, the crystal protein profile and plasmid DNA pattern of 2385-1 differed from those of subsp. kenyae. To verify the crystal protein gene types of 2385-1, a PCR-RFLP analysis was performed, and the results revealed that 2385-1 contained two novel cry1-type crystal protein genes, cryl-5 and cry1-12, in addition to the crylJal gene. The deduced amino acid sequences of cryl-5 and cry1-12 showed a 97.9% and 75.7% sequence similarity with the CrylAb and CrylJa crystal proteins, respectively. Among the novel crystal proteins, Cry1-5 showed a high toxicity towards P. xylostella and S. exigua larvae. In conclusion, B. thuringiensis 2385-1 is a new isolate in terms of its gene types, and should be a promising source for an insecticide to control lepidopteran larvae.

Comparative Analysis of Genetic Variation of Cucumber Mosaic Virus from Commelina communis in Korea (국내 닭의장풀에서 분리된 오이모자이크바이러스 분리주들의 외피단백질 유전자와 병징 다양성 비교)

  • Park, Tae-Seon;Hong, Jin-Sung
    • Research in Plant Disease
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    • v.24 no.2
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    • pp.170-173
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    • 2018
  • Three isolates of Cucumber mosaic virus isolated from Commelina communis plants showing chlorosis and mosaic were collected in Chungju and Chuncheon, Korea. To confirm genetic variation of these three isolates (CMV-Co, CMV-Co2, and CMV-Co3), we performed PCR-RFLP and sequence analysis. Sequences of coat protein genes of CMV-C0, -Co2 and -Co3 were compared with CMV-Fny and showed 96.3%, 96.3%, and 95.9% similarities, respectively. In host reactions, three CMV-Co isolates induced systemic necrosis in Cucurbita pepo unlike CMV-Fny and CMV-Co, CMV-Co2 and CMV-Co3 observed differential symptoms responses in Physalis angulata and Nicotiana rustica. These results indicated that three isolates of CMV isolated from C. communis have genetic and biological variation.

Sequence Variation in Superoxide Dismutase Gene of Toxoplasma gondii among Various Isolates from Different Hosts and Geographical Regions

  • Wang, Shuai;Cao, Aiping;Li, Xun;Zhao, Qunli;Liu, Yuan;Cong, Hua;He, Shenyi;Zhou, Huaiyu
    • Parasites, Hosts and Diseases
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    • v.53 no.3
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    • pp.253-258
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    • 2015
  • Toxoplasma gondii, an obligate intracellular protozoan parasite of the phylum Apicomplexa, can infect all warm-blooded vertebrates, including humans, livestock, and marine mammals. The aim of this study was to investigate whether superoxide dismutase (SOD) of T. gondii can be used as a new marker for genetic study or a potential vaccine candidate. The partial genome region of the SOD gene was amplified and sequenced from 10 different T. gondii isolates from different parts of the world, and all the sequences were examined by PCR-RFLP, sequence analysis, and phylogenetic reconstruction. The results showed that partial SOD gene sequences ranged from 1,702 bp to 1,712 bp and A + T contents varied from 50.1% to 51.1% among all examined isolates. Sequence alignment analysis identified total 43 variable nucleotide positions, and these results showed that 97.5% sequence similarity of SOD gene among all examined isolates. Phylogenetic analysis revealed that these SOD sequences were not an effective molecular marker for differential identification of T. gondii strains. The research demonstrated existence of low sequence variation in the SOD gene among T. gondii strains of different genotypes from different hosts and geographical regions.

Anisakis pegreffii Larvae in Sea Eels (Astroconger myriaster) from the South Sea, Republic of Korea

  • Cho, Jaeeun;Lim, Hyemi;Jung, Bong-Kwang;Shin, Eun-Hee;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • v.53 no.3
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    • pp.349-353
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    • 2015
  • Anisakis simplex sensu stricto (s.s.), Anisakis pegreffii, Anisakis berlandi (=A. simplex sp. C), and Anisakis typica are the 4 major species of Anisakis type I larvae. In the Republic of Korea (Korea), A. pegreffii, A. berlandi, and A. typica larvae in fish hosts has seldom been documented. In this study, molecular analysis was performed on Anisakis larvae from the sea eels (Astroconger myriaster), the major source of human anisakiasis in Korea, collected from Tongyeong City, a southern coastal area of Korea. All 20 sea eels examined were infected with Anisakis type I larvae (160 larvae; 8 per fish). Their species were analyzed using PCR-RFLP patterns and nucleotide sequences of internal transcribed spacers (ITS1, 5.8 subunit gene, and ITS2) and mitochondrial cytochrome c oxidase 2 (cox2). Most (86.8%; 112/129) of the Anisakis type I larvae were A. pegreffii, and 7.8% (10/129) were A. typica. The remaining 5.4% (7/129) was not identified. Thus, A. pegreffii is the major species of anisakid larvae in sea eels of the southern coast of Korea.

Prevalence of Toxoplasma gondii in Dogs in Zhanjiang, Southern China

  • Jiang, Hai-Hai;Li, Ming-Wei;Xu, Min-Jun;Cong, Wei;Zhu, Xing-Quan
    • Parasites, Hosts and Diseases
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    • v.53 no.4
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    • pp.493-496
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    • 2015
  • Toxoplasmosis, caused by Toxoplasma gondii, is a parasitic zoonosis with worldwide distribution. The present study investigated the prevalence of T. gondii in dogs in Zhanjiang city, southern China, using both serological and molecular detection. A total of 364 serum samples and 432 liver tissue samples were collected from the slaughter house between December 2012 and January 2013 and were examined for T. gondii IgG antibody by ELISA and T. gondii DNA by semi-nested PCR based on B1 gene, respectively. The overall seroprevalence of T. gondii IgG antibody was 51.9%, and T. gondii DNA was detected in 37 of 432 (8.6%) liver tissue samples. These positive DNA samples were analyzed by PCR-RFLP at 3'- and 5'-SAG2. Only 8 samples gave the PCR-RFLP data, and they were all classified as type I, which may suggest that the T. gondii isolates from dogs in Zhanjiang city may represent type I or type I variant. This study revealed the high prevalence of T. gondii infection in dogs in Zhanjiang city, southern China. Integrated measures should be taken to prevent and control toxoplasmosis in dogs in this area for public health concern.

Occurrence and Molecular Identification of Anisakis Dujardin, 1845 from Marine Fish in Southern Makassar Strait, Indonesia

  • Anshary, Hilal;Sriwulan, Sriwulan;Freeman, Mark A.;Ogawa, Kazuo
    • Parasites, Hosts and Diseases
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    • v.52 no.1
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    • pp.9-19
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    • 2014
  • Anisakis spp. (Nematoda: Anisakidae) parasitize a wide range of marine animals, mammals serving as the definitive host and different fish species as intermediate or paratenic hosts. In this study, 18 fish species were investigated for Anisakis infection. Katsuwonus pelamis, Euthynnus affinis, Caranx sp., and Auxis thazard were infected with high prevalence of Anisakis type I, while Cephalopholis cyanostigma and Rastrelliger kanagurta revealed low prevalence. The mean intensity of Anisakis larvae in K. pelamis and A. thazard was 49.7 and 5.6, respectively. A total of 73 Anisakis type I larvae collected from K. pelamis and A. thazard were all identified as Anisakis typica by PCR-RFLP analysis. Five specimens of Anisakis from K. pelamis and 15 specimens from A. thazard were sequenced using ITS1-5.8S-ITS2 region and 6 specimens from A. thazard and 4 specimens from K. pelamis were sequenced in mtDNA cox2 region. Alignments of the samples in the ITS region showed 2 patterns of nucleotides. The first pattern (genotype) of Anisakis from A. thazard had 100% similarity with adult A. typica from dolphins from USA, whereas the second genotype from A. thazard and K. pelamis had 4 base pairs different in ITS1 region with adult A. typica from USA. In the mtDNA cox2 regions, Anisakis type I specimens from A. thazard and K. pelamis showed similarity range from 94% to 99% with A. typica AB517571/DQ116427. The difference of 4 bp nucleotides in ITS1 regions and divergence into 2 subgroups in mtDNA cox2 indicating the existence of A. typica sibling species in the Makassar Strait.