• Title/Summary/Keyword: Gene Transfer

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Gene Expression Patterns of the Endogenous Antioxidant Enzymes in Linuron-Treated Rat Ventral Prostates after Castration

  • Yon, Jung-Min;Lin, Chunmei;Lee, Yoon-Bok;Lee, Beom-Jun;Yun, Young-Won;Nam, Sang-Yoon
    • Journal of Embryo Transfer
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    • v.27 no.2
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    • pp.101-105
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    • 2012
  • Linuron is a pesticide with a weak anti-androgenic property, which impacts male reproductive organs. In this study, to clarify whether linuron affects the cellular antioxidant system of ventral prostate, gene expression patterns of the representative antioxidant enzymes such as glutathione peroxidase (GPx), selenoprotein P (SePP), and superoxide dismutase (SOD) were investigated in the rat ventral prostates exposed to linuron using real-time RT-PCR analyses. Sprague-Dawley rats castrated at 6 weeks old were treated with linuron (25, 50, or 100 mg/kg per oral) daily for 10 days after testosterone propionate administration (0.4 mg/kg) subcutaneously. As compared to normal control animals, mRNA levels of phospholipid hydroperoxide GPx (PHGPx), SePP, and Mn SOD significantly increased in the prostates exposed to linuron (25, 50, and 100 mg/kg). However, cytosolic GPx (100 mg/kg) and Cu/Zn SOD (25, 50, and 100 mg/kg) mRNA levels significantly decreased in the ventral prostates. These results indicate that linuron upregulates the expressions of PHGPx, SePP, and Mn SOD mRNAs, but down-regulates the expressions of cytosolic GPx and Cu/Zn SOD in rat prostates, suggesting that linuron may have dual effects in the cellular antioxidant system of prostate.

Expression of PAPP-A and $20{\alpha}$-HSD in the Bovine Corpus Luteum during Early Pregnancy (소의 초기 임신 황체에서 PAPP-A와 $20{\alpha}$-HSD의 발현 양상)

  • Kim, Dae-Seung;Kim, Sang-Hwan;Yoon, Jong-Taek
    • Journal of Embryo Transfer
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    • v.26 no.1
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    • pp.57-63
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    • 2011
  • This study was performed to the expressions of pregnancy-associated plasma protein-A (PAPP-A) and 20alpha-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) in bovine corpus luteum during early pregnancy. To determine the function of PAPP-A gene during early pregnancy, we collected corpus luteum samples on 30, 60 and 90 days of pregnancy in bovine. The mRNA expression of PAPP-A, $20{\alpha}$-HSD, progesterone-receptor (PR) and insulin-like growth factor binding protein4 (IGFBP4) gene was conducted by Real-time PCR. In parallel with mRNA levels, The protein expressions of PAPP-A and $20{\alpha}$-HSD were detected by immunological analysis. The mRNA expressions $20{\alpha}$-HSD and PAPP-A significantly increased on day 90 in the corpus luteum during pregnancy. The mRNA expression of PR and JGFBP4 in the corpus luteum progressively was enhanced at 30 to 60 day, but decreased on 90 day of pregnancy in the corpus luteum. The expression patterns of these genes, PAPP-A and $20{\alpha}$-HSD were similar pattern in these tissues. In conclusion, PAPP-A and $20{\alpha}$-HSD activity in corpus luteum could be played a role for early pregnancy manifestation.

Effects of Genistein on the Gene Expressions of Glutathione Peroxidases and Superoxide Dismutases in Ethanol-Treated Mouse Fetuses

  • Yon, Jung-Min;Lin, Chunmei;Jung, A-Young;Lee, Jong-Geol;Jung, Ki-Youn;Baek, In-Jeoung;Lee, Beom-Jun;Nam, Sang-Yoon;Yun, Young-Won
    • Journal of Embryo Transfer
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    • v.26 no.2
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    • pp.135-140
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    • 2011
  • Genistein is a product of naturally occurring isoflavones at relatively high levels in soybeans. The harmful effects of ethanol are attributed to the induction of biological processes which lead to an increase in the generation of reactive oxygen species in fetuses. In this study, we investigated the effects of genistein ($1{\times}10^{-8}$ and $1{\times}10^{-7}\;{\mu}g$/ml) on gene expressions of the representative cellular antioxidative enzymes in ethanol (1 ${\mu}l$/ml)-treated mouse fetuses during the critical period (embryonic days 8.5~10.5) of organogenesis using a semi-quantitative RT-PCR analysis. The mRNA levels of cytosolic glutathione peroxidase (GPx), phospholipid hydroperoxide GPx, cytosolic CU,Zn-superoxide dismutase (SOD), and mitochondrial SOD were significantly decreased in ethanol-treated fetuses. However, the mRNA levels of ethanol plus genistein-treated fetuses were significantly higher than those of ethanol alone fetuses. These results indicate that genistein can up-regulate the expressions of GPx and SOD mRNAs reduced by the ethanol treatment in fetuses.

Identification of Pedigree by Bovine Blood Typing in Holstein (젖소에 있어서 혈액형분석에 의한 혈통확인에 관한 연구)

  • 이장희;신형두;정호영;유충원;안병석;이수헌;정상원;김창원
    • Journal of Embryo Transfer
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    • v.9 no.2
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    • pp.197-205
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    • 1994
  • This experiment was carried out to clarify the pedigree identification from blood typing of 301 Hoisteins in National Animal Breeding Institute(N.A.B.I.). Twenty kinds of standard reagent standardized by Insternational Society for Animal Blood Group Research provied from KNC improvement center, N, L, C, F. were used as the reference reagents in this study. The highest frequency of antigenic facfors was obtained from X$_2$in blood typing of 301 Holsteins. The frequency of X$_2$ was 0.714.In A blood system, four kinds of phenogroups were observed. The gene frequencies of Al and Z' phenogroups were equally 0.027.This frequency was greatly lower than those of breeds of Southern European and Zebu cattle. In B blood Systern, nineteen kinds of blood type were appeared. The appearance frequency of Gx blood type was 0.259, whish was higher than the others. In C blood system, thirty kinds of blood type were observed. The appearance frequency of X$_2$ blood type was the highest(0.189). In F blood system, three kinds of alleles were detected. The gene frequency of F allele was higher than that of V(0.105). However, the frequency of F allele(0.327) was greatly lower than that of "- /- " allele. In S blood system, twelve kinds of blood type were appeared and showed sirnilar appearance frequencies except " - / - " allele. From the results of the pedigree identification from 8 sires and 28 progenies of them, the accuracy of pedigree identification was 92.9%.ification was 92.9%.

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Evaluation of Three Candidate Genes Affecting Fatty Acid Composition in Pigs

  • Maharani, Dyah;Jung, Yeon-kuk;Jo, Cheorun;Jung, Woo-Young;Nam, Ki-Chang;Seo, Kang-Seok;Lee, Seung-Hwan;Lee, Jun-Heon
    • Food Science of Animal Resources
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    • v.32 no.1
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    • pp.6-12
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    • 2012
  • The association of three candidate genes, fatty acid synthase (FASN), microsomal triglyceride transfer protein (MTTP) and fatty acid binding protein 3 (FABP3), with fatty acid (FA) composition in Duroc pigs was investigated. Identified single nucleotide polymorphisms (SNPs) were used for polymerase chain reaction-restriction fragment length polymorphism genotyping. The c.265C>T SNP of FASN gene was significantly associated with high levels of palmitoleic acid (C16:1) (p<0.05), oleic acid (C18:1) (p<0.01), and mono-unsaturated fatty acid (MUFA) (p<0.01), but low levels of linoleic acid (C18:2) (p<0.01), alpha linolenic acid (C18:3) (p<0.05), and poly-unsaturated fatty acid (PUFA) (p<0.01) in animals having the CT genotype. The c.2573T>C SNP in the MTTP gene had a significant effect only in elevating the level of palmitoleic acid (C16:1) (p<0.05) in heterozygote animals. The polymorphism in FABP3 showed no significant effects on any fatty acid composition traits. These results suggest that the identified SNPs in the FASN and MTTP genes can be useful markers for selecting Duroc pigs having desirable healthy fatty acid composition.

Analysis of ROS and Apoptosis of Porcine Skin-derived Stem-like Cells after Differentiation Induction into Mesodermal Cell Types

  • Bae, Hyo-Kyung;Lee, Hwa-Yeon;Park, Yeo-Reum;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Journal of Embryo Transfer
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    • v.31 no.1
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    • pp.89-95
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    • 2016
  • The present study was conduct to examine the $H_2O_2$ expression level and apoptosis-related gene expression levels inporcineskin-derived stem cell-like cells (pSSCs) after adipogenic, chondrogenic, and osteogenic differentiation induction. The pSSCs were obtained by digestion of porcine ear skin biopsy and cultured in each induction medium for 21 to 26 days to induce adipogenic, chondrogenic, and osteogenic differentiation, respectively. The $H_2O_2$ levels of pSSCs after induction culture were evaluated by staining with 2'7'-dichlorodihydrofluorescein diacetate ($H_2DCFDA$). The apoptotic gene expression of pSSCs after induction culture was also estimated by RT-PCR. The pSSCs have a potential to differentiate into three mesodermal cell types (adipocytes, chondrocytes, and osteoblasts). Non-induced control and chondrogenic-induced cells were showed higher $H_2DCFDA$ intensity (P<0.05) than adipogenic- and osteogenic-induced cells. The relative expression of Bax/Bcl-2 level was significantly low (P<0.05) in adipogenic- and osteogenic-induced cells compared to non-induced control. However, there was no difference in the relative expression of Bax/Bcl-2 level among differentiation induction groups. The result of the present study shows that the apoptosis of pSSCs is not detrimentally increased by differentiation induction culture, although chondrogenic-induced pSSCs showed high ROS generation level and apoptotic index similarly to those of non-induced cells.

An Analysis of Survey Data on South Korea Boar Stud Practices (한국의 돼지 인공수정센터 현황 분석)

  • Sa, Soo-Jin;Woo, Jae-Seok;Hong, Joon-Ki;Kim, Ki-Hyun;Kim, Du-Wan;Kim, Yong-Min;Park, Hyun-Sik;Kim, Si-Joo;Chung, Ki-Hwa;Cho, Eun-Seok
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.129-135
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    • 2015
  • The aim of this study is to examine current status of swine AI and boar stud in South Korea using survey and data analysis. This survey included 48 boar studs registered as 'semen processing business'. The survey data were collected by direct visitation, FAX and/or telephone conversation for 7 months from June through December in 2013. 48 boar studs owned a total of 3,537 boars and the Duroc breed accounted for the highest rate (75.3%) of all boar breeds. In case of ownership, agricultural management corporations was the highest (50.0%) and followed by individual ownership (33.3%). Large-scale boar studs in terms of own over 151 boar were surveyed as 4.2% and most boar studs owned less than 100 boars (77.1%). The amount of liquid semen provided by 48 boar studs were 1,889,000 doses and each boar stud provided average of 39,000 does, which is represented for 90% consumption by sows in South Korea.

Confirmation of Male Specific Fetal Free RNA in Maternal Plasma and Comparison of Accuracy on the Sex Determination using Real-time PCR Method in Korean Native Cattle

  • Lee, Sang-Ho;Park, Chul-Ho;Park, Jun-Tae;Park, Sang-Guk;Lee, Jin-A;Suh, Guk-Hyun;Oh, Ki-Seok;Son, Chang-Ho
    • Journal of Embryo Transfer
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    • v.28 no.4
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    • pp.343-348
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    • 2013
  • Cell-free fetal RNA has been highlighted as useful tools for the fetal sex determination or other genetic inherent disorder. However, there is no knowledge about the sex determination using cell free fetal RNA in bovine field. Thus, the present study aimed to evaluate the presence of transcripts of DDX3Y, USP9Y and ZRSR2Y genes in maternal plasma of pregnant cows to determine the sex of the fetus using real-time quantitative polymerase chain reaction assay, and verify its accuracy, sensitivity and specificity compared with the molecular testing and the calf sex at birth. Transcripts of USP9Y and DDX3Y genes were expressed in the all plasma of males and females both the control group and the experimental group. However, ZRSR2Y gene was matched up with the molecular testing and the true sex in control group and has an overall accuracy of 82.6%, a sensitivity of 75%, and a specificity of 100% in experimental group. Therefore, these results indicated that real time PCR technique, as a noninvasive and cost-efficient method, is possible to determination fetal sex in the bovine species using circulating cell free RNA in maternal plasma and especially ZRSR2Y gene could be a good candidate for the RNA based sex determination work.

Expression and evaluation of porcine circovirus type 2 capsid protein mediated by recombinant adeno-associated virus 8

  • Li, Shuang;Wang, Bo;Jiang, Shun;Lan, Xiaohui;Qiao, Yongbo;Nie, Jiaojiao;Yin, Yuhe;Shi, Yuhua;Kong, Wei;Shan, Yaming
    • Journal of Veterinary Science
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    • v.22 no.1
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    • pp.8.1-8.11
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    • 2021
  • Background: Porcine circovirus type 2 (PCV2) is an important infectious pathogen implicated in porcine circovirus-associated diseases (PCVAD), which has caused significant economic losses in the pig industry worldwide. Objectives: A suitable viral vector-mediated gene transfer platform for the expression of the capsid protein (Cap) is an attractive strategy. Methods: In the present study, a recombinant adeno-associated virus 8 (rAAV8) vector was constructed to encode Cap (Cap-rAAV) in vitro and in vivo after gene transfer. Results: The obtained results showed that Cap could be expressed in HEK293T cells and BABL/c mice. The results of lymphocytes proliferative, as well as immunoglobulin G (IgG) 2a and interferon-γ showed strong cellular immune responses induced by Cap-rAAV. The enzyme-linked immunosorbent assay titers obtained and the IgG1 and interleukin-4 levels showed that humoral immune responses were also induced by Cap-rAAV. Altogether, these results demonstrated that the rAAV8 vaccine Cap-rAAV can induce strong cellular and humoral immune responses, indicating a potential rAAV8 vaccine against PCV2. Conclusions: The injection of rAAV8 encoding PCV2 Cap genes into muscle tissue can ensure long-term, continuous, and systemic expression.

Current status on the molecular biological research for the origin of cultivated sweetpotato [Ipomoea batatas L. (Lam)] (재배종 고구마[Ipomoea batatas L. (Lam)]의 기원종에 관한 분자생물학적 연구 동향)

  • Lee, Shin Woo;Kim, Yun-Hee
    • Journal of Plant Biotechnology
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    • v.48 no.4
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    • pp.223-227
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    • 2021
  • Several hypotheses for the origin of cultivated sweetpotato [Ipomoea batatas L. (Lam)] have been suggested but the exact progenitor is still unknown. Based on the results of RFLP patterns, microsatellite markers, SNP markers, FISH analyses, and genome analyses of haplotypes, wild species belonging to batatas group, I. trifida, I. leucantha, I. littoralis, I. tabascana, I. tenuissima, I. tiliacea, and I. triloba have been suggested as a progenitor. However, recently, advanced genomic technologies and characterization of the inserted T-DNA fragments of Agrobacterium in the genome of cultivated sweetpotato and wild species through horizontal gene transfer suggest that there may be an older progenitor than the wild species suggested so far.