• Title/Summary/Keyword: Cloning efficiency

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Effects of variation in the number and developmental stage of donor embryos and ovulation status of the surrogate mother on the efficiency of pig somatic cell cloning

  • Park, Mi-Ryung;Yoo, Jae Gyu;Hur, Chang-Gi;Sim, Bo-Woong;Kim, Myunghoo;Seo, Jakyeom;Kim, Byeong-Woo;Cho, Byung-Wook;Shin, Teak-Soon;Cho, Seong-Keun
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.3
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    • pp.258-264
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    • 2020
  • This study investigated the effect of variation in the number of somatic-cell-cloned embryos and their developmental stage at transfer on pregnancy, as well as the influence of the estrus status of recipient pigs on in vivo development of cloned porcine embryos after embryo transfer. For somatic cell nuclear transfer (SCNT), fibroblast cells were obtained from a male porcine fetus. Recipient oocytes were collected from prepubertal gilts at a local abattoir and then cultured. After SCNT, reconstructed embryos of different numbers and developmental stages were transferred into recipient pigs. The developmental stage of the cloned embryos and the number of transferred embryos per surrogate showed no significant differences in terms of the resulting cloning efficiency. However, the pregnancy rate improved gradually as the number of transferred cloned embryos was increased from 100-150 or 151-200 to 201-300 per recipient. In pre-, peri-, and post-ovulation stages, pregnancy rates of 28.6%, 41.8%, and 67.6% and 16, 52, and 74 offspring were recorded, respectively. The number of cloned embryos and estrus status of the recipient pig at the time of transfer of the cloned embryo affect the efficiency of pig production; therefore, these variables should be particularly considered in order to increase the efficiency of somatic cell pig cloning.

Map based cloning of resistance to bacterial leaf blight gene using QTL analysis in rice

  • Du, Xiao-Xuan;Kim, Kyung-Min
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.138-138
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    • 2017
  • Agriculture is the most primitive civilized Activities of mankind but also the propellant of civilization development. Because it is the most basic material goods source of mankind. Among these materials rice is one of the most important part of these, we call them the substance of survival. From the beginning of the agricultural activities to the present we have experienced three industrial revolutions and are experiencing the Fourth Industrial Revolution. With the development of science and technology makes the efficiency of agricultural production is higher and higher, but compared with the original we are facing the same problem: natural disasters; pests and diseases; now also face the depletion of resources, environmental degradation and other issues. Therefore, improve and cultivate new crop varieties to make it better resistance and more production for better develop modern agriculture. It's very helpful for human social development. And also it is the responsibility and task of modern molecular breeding. In this study, I used bacterial leaf blight to find a better resistance gene to improve the resistance of rice. Frist Cultivate k3 of bacterial leaf blight, than inoculation by leaf clipping method (Kauffman,1973) in CNDH and SNDH population at 40days after rice transplanting. Check the lesion length by inoculation plants at 14days after inoculation, and record data for QTL analysis program. Than I get 4 intervals in 3 different chromosomal regions. I found these defense genes in the 4 intervals. So I used NCBI Justbio, Rapdb, etc. to finding these genes in physical map, than design primer for map base cloning. At last these defense genes will be employed in further research for introduction of the gene to the parental plant and rice breeding for solving food crisis.

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The Effects of Resveratrol on Oocyte Maturation and Preimplantation Embryo Development

  • Kwak, Seong-Sung;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.27 no.2
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    • pp.71-80
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    • 2012
  • Biotechnologies for cloning animals and in vitro embryo production have the potential to produce biomedical models for various researches. Especially, pigs are a suitable model for xenotransplantation, transgenic production and various areas of reproductive research due to its physiological similarities to human. However, utilization of in vitro-produced embryos for transfer remains limited. Despite improvement over past few decades, obstacles associated with the production of good quality embryos in vitro still exist which limit the efficiency of cloning. One of major problems includes improper in vitro maturation (IVM) and culture (IVC). Oxidative stress caused from in vitro culture conditions contributes to inadequate IVM and IVC which leads to poor developmental competence of oocytes, failure of fertilization and embryo development. To reduce the oxidative stress, various antioxidants have been used to IVM and IVC system. However, limited information is available on the effects of resveratrol on livestock reproductions. Resveratrol is a polyphenolic natural product and well known as an antioxidant in foods and beverages (e.g. in grapes and red wine). Resveratrol is known to be cardioprotective, anticarcinogenic, anti-inflammatory, antioxidant and antiapoptotic. This paper will review the effects of resveratrol on in vitro maturation of oocytes and embryo development.

Cloning and Expression of D-amino Acid Oxidise from Trigonopsis variabilis for Cephalosporin C Biotransformation (Cephalosporin C의 생변환을 위한 Trigonopsis variabilis의 D-amino Acid Oxidase 유전자의 클로닝 및 발현)

  • 이진형;정태완
    • KSBB Journal
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    • v.10 no.3
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    • pp.264-270
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    • 1995
  • Trigonopsis variabilis is a strong producer of D-amino acid oxidase that can transform cephalosporin C(ceph C) to ${\alpha}$-keto-adipyl-7-aminocephalosporanic acid(AKA-7ACA). Polymerase chain reaction (PCR) was applied to isolate the D-AAO gene from T. variabilis. To clone the PCR fragment, four different methods were examined using enzymatic reactions of Taq DNA polymerase, Klenow, T4 DNA polymerase I, Alkaline phosphatase Calf Intestinal, and T4 kinase. Ligation of phosphorylated blunt-end PCR fragment and dephosphorylated blunt-end of pUC18 plasmid yielded the best cloning efficiency One of recombinant E. coli transformants showed D-AAO activity against ceph C in both cell extracts and permeabilized cells.

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Cloning and Iron Transportation of Nucleotide Binding Domain of Cryptosporidium andersoni ATP-Binding Cassette (CaABC) Gene

  • Wang, Ju-Hua;Xue, Xiu-Heng;Zhou, Jie;Fan, Cai-Yun;Xie, Qian-Qian;Wang, Pan
    • Parasites, Hosts and Diseases
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    • v.53 no.3
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    • pp.335-339
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    • 2015
  • Cryptosporidium andersoni ATP-binding cassette (CaABC) is an important membrane protein involved in substrate transport across the membrane. In this research, the nucleotide binding domain (NBD) of CaABC gene was amplified by PCR, and the eukaryotic expression vector of pEGFP-C1-CaNBD was reconstructed. Then, the recombinant plasmid of pEGFP-C1-CaNBD was transformed into the mouse intestinal epithelial cells (IECs) to study the iron transportation function of CaABC. The results indicated that NBD region of CaABC gene can significantly elevate the transport efficiency of $Ca^{2+}$, $Mg^{2+}$, $K^+$, and $HCO_3{^-}$ in IECs (P<0.05). The significance of this study is to find the ATPase inhibitors for NBD region of CaABC gene and to inhibit ATP binding and nutrient transport of CaABC transporter. Thus, C. andersoni will be killed by inhibition of nutrient uptake. This will open up a new way for treatment of cryptosporidiosis.

Production of Cloned Korean Native Pig by Somatic Cell Nuclear Transfer

  • Hwang, In-Sul;Kwon, Dae-Jin;Oh, Keun Bong;Ock, Sun-A;Chung, Hak-Jae;Cho, In-Cheol;Lee, Jeong-Woong;Im, Gi-Sun;Hwang, Seongsoo
    • Development and Reproduction
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    • v.19 no.2
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    • pp.79-84
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    • 2015
  • The Korean native pig (KNP) have been considered as animal models for animal biotechnology research because of their relatively small body size and their presumably highly inbred status due to the closed breeding program. However, little is reported about the use of KNP for animal biotechnology researches. This study was performed to establish the somatic cell nuclear transfer (SCNT) protocol for the production of swine leukocyte antigens (SLA) homotype-defined SCNT KNP. The ear fibroblast cells originated from KNP were cultured and used as donor cell. After thawing, the donor cells were cultured for 1 hour with 15 ${\mu}M$ roscovitine prior to the nuclear transfer. The numbers of reconstructed and parthenogenetic embryos transferred were $98{\pm}35.2$ and $145{\pm}11.2$, respectively. The pregnancy and delivery rate were 3/5 (60%) and 2/5 (40%). One healthy SLA homotype-defined SCNT KNP was successfully generated. The recipient-based individual cloning efficiency ranged from 0.65 to 1.08%. Taken together, it can be postulated that the methodological establishment of the production of SLA homotype-defined cloned KNP can be applied to the generation of transgenic cloned KNP as model animals for human disease and xenotransplantation researches.

Efficiency of Female-Derived Donor Cells on High Postnatal Survival in Pig Cloning

  • Cho, Seong-Keun;Park, Mi-Rung;Kwon, Deug-Nam;Hwang, Kyu-Chan;Lee, Eun-Kyeong;Son, Woo-Jin;Kim, Jin-Hoi
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.217-217
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    • 2004
  • The present study was conducted to investigate the developmental competency between male- and female-somatic cell derived nuclear-transferred porcine embryos, and the productive and survival efficiency of cloned male and female piglets. The potential of eggs receiving somatic cells to develop into blastocysts was not different among donor cells of different origins. (omitted)

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Effect of DO Concentration on Ammonia Oxidizing Bacteria in Aerobic Biofilm Reactor (호기성 생물막 반응기에서 Ammonia Oxidizing Bacteria에 대한 DO 농도의 영향)

  • Yu, Jae-Cheul;Park, Jeung-Jin;Hur, Sung-Ho;Kim, Yu-Jin;Byun, Im-Gyu;Lee, Tae-Ho;Park, Tae-Joo
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.1
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    • pp.106-112
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    • 2007
  • Ammonia oxidizing bacteria(AOB) oxidize ammonia to nitrite and are important microorganisms which control nitrification. Several environmental factors such as dissolved oxygen(DO), temperature and pH influence the growth of AOB. In this work, to assess the effect of DO concentration on AOB, four aerobic biofilm reactors packed with ceramic media were operated 1, 3, 5 and 7 mgDO/L, respectively. The optimal DO concentration with stable nitrification efficiency in aerobic biofilm reactor was above 5.0 mg/L. To assess the relationship between the DO concentration and the characteristics of AOB in aerobic biofiim reactor, DGGE and cloning based on PCR targeting 16S rRNA and amoA gene were performed. Additionally, INT-DHA activity test was proceeded to estimate the activity of AOB. As the results of DGCE and cloning, the community of AOB and the ratio of Nitrosomonas sp. changed little in spite of different nitrification efficiencies. INT-DHA activity test showed that the activity of AOB decreased as DO concentration decreased. It means that DO concentration does not affect the community of AOB, but the activity of AOB.

Effect of Non-homologous Spacing in Target DNA Sequence on the Frequency of Cloning Based Homologous Recombination (Target DNA 염기서열 내에 존재하는 비상동성 간격이 상동성재조합을 이용한 클로닝 빈도에 미치는 영향)

  • Kim Jae-Woo;Do Eun-Ju;Yoon Se-Lyun;Jeong Yun-Hee;Yoon Young-Ho;Leem Sun-Hee;Sunwoo Yangil;Park In-Ho
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.239-245
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    • 2005
  • Transformation-Associated Recombination (TAR) cloning technique allows selective isolation of chromosomal regions and genes from complex genomes. The procedure requires knowledge of relatively small genomic sequences that reside adjacent to the chromosomal region of interest. This technique involves homologous recombination during yeast spheroplast transformation between genomic DNA and a TAR vector that has 5' and 3' gene targeting sequences. In this study, we examined the effect of non-homologous spacing sequence in target hooks on homologous recombination using a plasmid model system. The efficiency of homologous recombination between the modified his3-TRP1-his3 fragments and HlS3 gene on plasmid were analyzed by the characterization of $Ura^+$ transformants. The numbers of $Ura^+$ transformant showed same level when seven different modified his3-TRP1-his3 fragments were used. But the percentage of positive recombinants. $Trp^+His^-$, dramatically decreased when used the modified his3-TRP1-his3 fragments contained incorrect spacing of nonhomologous region. As a result, we suggest that incorrect spacing inhibits the homologous recombination between target hook and substrate DNA. Therefore, we should consider the correct spacing in target hook when the target hook are used for cloning of orthologue gene.

e-Passport Integrated Authentication Mechanisms with Improved Efficiency (효율성이 강화된 전자여권 통합 인증 메커니즘)

  • Lee, Dong-Bum;Go, Woong;Kwak, Jin
    • Journal of Internet Computing and Services
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    • v.10 no.2
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    • pp.53-63
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    • 2009
  • e-Passport system is new type of emigration and immigration control system and it is a research to introduce the e-Passport Authentication Protocol with Improved Efficiency is lively proceeded over the entire world. The e-Passport's chip has a biometric information and personal identification information, Radio Frequency Identification(RFID) technology is used for communication with the Inspection System(IS). However, the feature of the RFID system may bring about various security threats such as eavesdropping, data forgeries, data alternation, cloning, biometric data-leakage. Therefore, in this paper, we analyse the e-Passport system's authentication protocol to protect vulnerability and proposed e-Passport system's authentication protocol reduce computation. Also, we compared their efficiency.

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