• 제목/요약/키워드: Cloning efficiency

검색결과 90건 처리시간 0.026초

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
    • 한국동물생명공학회지
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    • 제35권3호
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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
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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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
    • 한국수정란이식학회지
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    • 제27권2호
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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.

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

  • 이진형;정태완
    • KSBB Journal
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    • 제10권3호
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    • pp.264-270
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    • 1995
  • Trigonopsis variabilis는 버l타락탐 항생제인 cephalosporin C (ceph C)를 ${\alpha}$-keto-adipyl-7 a aminocephalosporanic acid(AKA-7 ACA)로 생변 환하는 강력한 D-amino acid oxidase 효소를 갖고 있다. 본 연구는 이 D-AAO 효소의 유전자를 추출하기 위하여 polymerase chain reaction (PCR)을 사용하였다. PCR 단편을 콜로닝하기 위하여 Taq D DNA polymerase, Klenow, T4 DNA polymerase I, Alkaline phosphatase Calf Intestinal와 T4 kinase 의 효소반응을 이용하여 4가지의 방법을 샤용한 결 과, blunt - end 의 PCR fragment 를 phosphory­l lation하고 blunt -end의 pUC18 plasmid를 dephos phorylation 한 후 ligation 한 것 이 가장 좋은 클로 닝 효율을 보였다. Ceph C에 대한 D-AAO 효소의 활성은 재조합 E. coli의 세포추출액과 permea bilized cells에서 모두 확인할 수 있었다.

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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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    • 제53권3호
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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
    • 한국발생생물학회지:발생과생식
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    • 제19권2호
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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
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2004년도 춘계학술발표대회
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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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호기성 생물막 반응기에서 Ammonia Oxidizing Bacteria에 대한 DO 농도의 영향 (Effect of DO Concentration on Ammonia Oxidizing Bacteria in Aerobic Biofilm Reactor)

  • 유재철;박정진;허성호;김유진;변임규;이태호;박태주
    • 대한환경공학회지
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    • 제29권1호
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    • pp.106-112
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    • 2007
  • Ammonia oxidizing bacteria(AOB)는 $NH_4^+-N$$NO_2^--N$으로 산화시키며, 생물학적 질산화 단계에서 율속 단계로 작용하기 때문에 중요한 미생물이다. AOB의 성장은 용존산소, 온도, pH 등의 환경 인자에 영향을 받는다. 본 연구에서는 DO 농도가 AOB에 미치는 영향을 조사하기 위해 세라믹 메디아가 충전된 4개의 호기성 생물막 반응기의 DO 농도를 각각 1, 3, 5, 7 mg/L로 운전하였다. 운전결과, 5 mg/L 이상에서 안정적인 질산화 효율을 얻을 수 있었다. AOB의 특성을 조사하기 위해 AOB의 16S rRNA와 amoA gene을 target으로 PCR을 이용한 DGGE와 cloning을 실시하였으며, 이들의 활성을 조사하기 위해 INT-DHA를 측정하였다. DO 농도 변화에 따른 각 반응기별 질산화 효율에 차이가 있었음에도 불구하고, DGGE 및 cloning 결과, AOB 군집 및 Nitrosomonas sp.의 비율의 변화는 거의 없었다. DO 농도가 감소함에 따라 AOB의 활성도가 감소한다는 것을 INT-DHA 측정으로 확인할 수 있었다. 따라서 DO 농도는 AOB 군집의 변화 보다는 활성에 영향을 미치는 것으로 판단되었다.

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

  • 김재우;도은주;윤세련;정윤희;윤영호;임선희;선우양일;박인호
    • 미생물학회지
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    • 제41권4호
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    • pp.239-245
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    • 2005
  • Transformation-associated recombination (TAR) 클로닝 법은 복잡한 게놈으로부터 염색체 내의 특정부위나 유전자를 선택적으로 분리할 수 있다. 이 방법은 목적 유전자에 근접한 작은 게놈DNA 염기서열 정보를 필요로 한다. 이 기술은 효모의 spheroplast transformation을 시키는 동안 목적으로 하는 유전자의 5' 또는 3' 서열을 포함하고 있는 TAR vector와 게놈DNA사이에서 일어나는 상동성재조합에 의해 이루어진다. 본 연구에서는 plasmid 모델시스템을 이용하여 target hooks 내에 존재하는 비상동성 염기서 열이 상동성재조합에 미치는 영향을 조사하였다. plasmid에 존재하는HIS3유전자와 변형시킨 his3-TRP1-his3 단편 사이의 상동성재조합의 효율은 $Ura^+$ 형질전환체의 형질분석에 의해 이루어졌다. $Ura^+$ 형질전환체의 수는 7종류의 서로 달리 변형된 his3-TRP1-his3 단편들을 사용하였을 매 거의 동일하게 나타났다. 그러나 $Trp^+His^+$ positive recombinants의 빈도는 변형된 his3-TRP1-his3 단편 내에 비상동성 영역에 부정확한 간격을 지닐 때 현저한 감소를 나타내었다. 이러한 결과로서, 부정확한 간격이 target hook과 substrate DNA 사이에 일어나는 상동성재조합을 방해하는 것으로 사료된다. 그러므로 이종간의 상동유전자를 클로닝 할 때에는 target hook내의 비상동성 염기서열이 존재한다면 이것이 정확한 간격을 지니는지 여부를 중요란 요인으로 고려해야 한다.

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

  • 이동범;고웅;곽진
    • 인터넷정보학회논문지
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    • 제10권2호
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    • pp.53-63
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    • 2009
  • 전자여권(e-Passport) 시스템은 새로운 형태의 출입국 관리 시스템으로서, 기존 여권보다 보안 기능이 강화되고 자동화된 출입국 관리를 할 수 있다는 장점으로 인해 전 세계적으로 도입하기 위한 연구가 활발히 진행되고 있다. 전자여권은 칩에 개인 신원 정보와 바이오정보를 저장하고, 비접촉식 무선 인식 기술을 이용하여 판독시스템과 통신을 제공하게 된다. 하지만 기존의 RFID 기술에서 발생하는 도청, 데이터 위 변조, 복제와 같은 문제점과 개인의 고유한 바이오정보 노출이라는 문제점을 내재하고 있다. 따라서 본 논문에서는 보안 취약점을 방지하는 전자여권 인증 메커니즘들에 대한 보안 취약점을 분석하고 계산량을 감소시킬 수 있는 전자여권 인증 메커니즘을 제안한다. 또한 각 인증 메커니즘의 효율성에 대하여 분석한다.

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