• Title/Summary/Keyword: 미세주입

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Production of Transgenic Pig Harboring the Cellulase Digest Gene(CelD) (섬유소 분해효소 유전자가 도입된 형질전환 돼지 생산)

  • 박진기;이연근;민관식;이창현;이향흔;김광식;장원경;김진회;이훈택
    • Korean Journal of Animal Reproduction
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    • v.26 no.2
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    • pp.87-94
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    • 2002
  • This study was performed during the four seasons for the production of transgenic pigs containing the Cellulase Digest Gene. Purebred Landrace gilts and sows approximately 8∼15 months of age (n=126) were used for the collection of 1-cell zygotes for DNA microinjection and transfer. Retrospectively, estrus synchronization and superovulation schemes were evaluated to assess practicality fur zygote collection. Synchronization and superovulation procedures were used that cyclic gilts were synchronized with 20mg altrenogest (ALT) per day for 9 days after PG600 administration followed by superovulation with 1000 IU pregnant mares serum gonadotropin (PMSG) and 750IU human chorionic gonadotrophin (hCG). The cellulase digestion gene for microinjection is rat elasterase promoter (rEl) linked to CelD gene. After hormone treatment, 1,422 embryos were collected from 91 donors and 95.6% (1,359/1,422) embryos were in 1-cell stage which can be visualized the pronuclei for DNA microinjection. A total of 725 DNA microinjected embryos transferred into 35 recipients and produced 65 piglets from 13 litters. Pregnancy rate according to the number of transferred embryos to recipients was higher the group which received 21 to 24 embryos (50.0%) than other groups 20.0% in less and 33.3% in more. A tail tissue was collected from 65 piglets for biopsy. PCR screening was performed on each DNA sample using two separate sets of primers specific for the 5'- and 3'-flanking region of the rEl-CelD gene. Five of the 65 piglets (7.69%) were positive for the transgene. This study provide useful information regarding production of transgenic pig for bioreactor research.

Studies of Micro-Air Flotation for Removal of Turbidity (탁도제거를 위한 미세공기 부양법 연구)

  • Choi, Boram;Kim, Dongsoo;Kim, Jongoh;Kim, Taeyoon
    • Journal of the Korean GEO-environmental Society
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    • v.14 no.8
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    • pp.23-27
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    • 2013
  • In this study, efficiency of pre-treatment of turbid seawater was measured where micro-air bubbles were used to remove particles in seawater after input of natural coagulant PGA. Artificial seawater was prepared having the intended trubidity using marine sediments and microalgae. 73.7% of turbidity removal was achieved when 0.5g/L of $AlCl_3{\cdot}6H_2O$ was added in the artificial seawater, but 92.4% of turbidity removal was observed when 0.05g/L of PGA was added in the artificial seawater containing microalgae. In addition, much greater turbidity removal was achieved for microalage than sediments. For both cases, input of 0.1g/L PGA and following additional input of micro-air bubbles for 5 seconds resulted in the maximum removal efficiency where reaction time of coagulation was 1 min and flotation by micro-air bubbles was 10 min. From this study, we concluded that micro-air floation after coagulation could be a possible economical pre-treatment method for highly turbid seawater.

In Vitro Development of Porcine Oocytes Following Intracytoplasmic Injection of Round Spermatid (원형정자 미세주입에 의한 돼지 난자의 체외 배 발달)

  • Shin J. S.;Kim, C.;S. J. Uhm;Kim, N. H.;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.395-403
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    • 1998
  • In this study we determined fertilization processes and developmental ability of porcine oocytes following injection of round spermatid in the presence and absence of artificial activation. Electrical stimulation at 2 h before spermatid injection significantly increased the incidence of normal fertilization as compared to those following injection without stimulation or with stimulation immediately after injection. The incidences of two pronuclear formation and apposition were not different in oocytes between following intracytoplasmic spermatid and spermatid nucleus injection. Indirect immunocyto chemistry and laser scanning confocal microscopy study revealed that microtubuels were organized from the oocyte cortex following round spermatid injection, and this seemed to move both male and female pronuclei into the oocyte center. At 6 days following injection blastocoele formation was seen in the eggs following round spermatid (25%) and round spermatid nucleus injection (27%). However, none of oocytes developed to the blastocyst stage at 6 days following sham injection. The average cell numbers of blastocysts at 8 days following injection of spermatid and spermatid nucleus were 87 to 99. These results suggested that either round spermatid or it's nudeus can be used to produce viable embryos by injection into unfertilized oocytes in the pig.

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A Study of the Liposome-Mediated Transgenic Chicken Production (리포좀을 이용한 형질전환 닭 생산에 대한 연구)

  • Byun S. J.;Park C.;Yang B. S.;Kim T. Y.;Sohn S. H.;Kim S. H.;Jeon I. S.
    • Korean Journal of Poultry Science
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    • v.31 no.4
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    • pp.293-298
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    • 2004
  • Microinjection of DNA is a general method for generating transgenic animals, but the rate of transgenesis in chickens is very low. So it was carried out to investigate the efficiency of liposome-mediated gene transfer in stage one cell of chicken embryo with GFP expression vector. In order to determine efficiency and duration of the introduced foreign gene, it was microinjected DNA with liposome or naked DNA into the germinal disc of stage one cell or stage-X chicken embryo. Analysis of reporter gene expression in day-4 embryos showed that GFP expression was observed only in the liposome-mediate embryo groups and detectable up to day-8 embryos. The results suggest that stable integration of the introduced gene using liposome is a rare event. Nevertheless the liposome-mediated gene transfer may be a useful method to transfer a foreign gene into the stage one cell of chicken embryos.

Effects of Sucrose Treatment on the Morphology and Integration of foreign DNA into Bovine Oocytes (소 난자에서 형태와 외래 DNA Integration에 관한 Sucrose 처리의 효과)

  • Kim, S. G.;Kim, K. S.;Kim, T. W.;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.25 no.4
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    • pp.399-407
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    • 2001
  • The microinjection of retroviral vectors into the perivitelline spaces of MII-stage oocytes increased production of transgenic bovine embryos. However, oocytes have various sizes of perivitelline space, and there is the tendency that the oocyte membranes are damageable by micropipettes during the injection of retroviral vectors into perivitelline spaces or oocytes. Thus, it was not always possible to stably inject retroviral vector into perivitelline spaces of oocytes. Here we used sucrose to minimize the damage of the oocyte membrane. When the oocytes were suspended in 0.5% sucrose, poor quality oocytes showed rough cytoplasmic membranes, while good quality oocytes maintained smooth membranes. However, when the tatters were subjected to in vitro fertilization, no significant differences were observed in cleavage rates (82% of control Vs. 84% of sucrose treated oocytes). The Same trends were obtained from the oocytes fertilized after microinjection of LN$\beta$-EGFP and LNC-hGH genes into the perivitelline spares. The rates of cleavage and blastocyst from microinjection of LN$\beta$-EGFP genes were 81 and 25%, and from microinjection of LNC-hGM genes were 53 and 30%, respectively. The result indicated that microinjected oocytes could develop to the blastocyst stages after in vitro fertilization with no significant difference from control group. Moreover, the integration of hGH-gene (by PCR analysis) was detected in 52% of infected cleaved embryos and the expression of EGFP-gene (under a fluoresrence microscope) was also observed in 34% of infected blastocyst. These results indicated that 0.5% sucrose treatment could be an efficient method not only to select good quality embryos but also to inject retroviral vectors into perivitelline spares without any harm and hence improving developmental rates.

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Comparison of Viable Rates of Chick Embryos by Different Eggshell Window Positioning (닭 배자 조작을 위한 난각 주입부위별 생존율 비교)

  • J. Y. Han;D. S. Seo;Y. H. Hong;D. K. Jeong;Y. S. Shin
    • Korean Journal of Poultry Science
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    • v.23 no.1
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    • pp.9-17
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    • 1996
  • This study was conducted to compare the survival rate of chick embryos among different eggshell window positions and to search for the most appropriate injection position. The eggshells were punctured at blunt-end, sharp-end and side-up with a sterilized fine forceps, respectively. The survival rate of sharp-end window was higher than the other window positions. Injection of Dulbecco’s modified eagle’s medium (DMEM) through blunt-end window (BE1) was impossible because inner cell membrane was obscure. The 2 ${\mu}$L DMEM was injected into 2.5 d-old embryo blood vessel through sharp end window. To prevent hemorrhages at the point of injection, the air bubbles were injected into the embryo blood vessel. The survival rate of chicks embryo in sharp end window was about 17.0%. Therefore, this sharp-end window system will be helpful for the production of germline chimera or transgenic chicken using primordial germ cells ( PGCs ).

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Fine Flow Controlling Device for Medicine Injection (의료 약물주입용 미세 유량 제어 장치)

  • Cho, Su-Chan;Shin, Bo-Sung
    • Journal of the Microelectronics and Packaging Society
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    • v.28 no.4
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    • pp.51-55
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    • 2021
  • The nurses manually carry out the intravenous therapy for the patients. Using an Arduino, the fine flow controlling device was invented to provide an ongoing patient care. The medication is injected through a peristaltic pump, and the amount of the solution is controlled with a RGB color sensor. The power of the device is supplied through the batteries. An amount of the injection is measured with LIG strain sensor fabricated by 355nm UV pulsed laser. This system will provide a better medical service.

Hydrogen Delayed Fracture of Automotive Multi-Phase Sheet Steels by Hydrogen Charging Condition (수소주입에 따른 자동차용 복합조직강판의 지연파괴 거동)

  • Park, Jae-U;Gang, Gye-Myeong
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2013.05a
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    • pp.87-87
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    • 2013
  • 복합조직강의 미세조직제어를 통한 자동차용 고강도 강판재의 개발이 주요 연구과제로 있다. 하지만 고강도화에 따라 수소에 의한 지연파괴의 문제점이 있어, 이를 규명하고 해결하기 위한 많은 연구가 함께 이루어지고 있다. 본 연구에서는 연구, 개발되고 있는 복합조직강 중 DP강과 TRIP강의 수소취성에 미치는 미세조직의 영향을 분석하고, 수소주입조건에 따른 수소취성 및 지연파괴 거동에 대하여 조사하고자 하였다. 이를 위해 음극전기분해법을 이용, 주입수소량을 달리하여 주입수소가 복합조직강의 지연파괴에 미치는 영향을 분석하였다. Hydrogen determinator를 통해 시편 내 수소량을 측정하였고, 소형펀치시험에 의한 기계적물성 변화를 조사하였다. 또한 파단부위의 넓이와 깊이를 비교측정하였고, 파단면을 SEM으로 관찰하여 수소지연파괴 거동을 평가하고자 하였다.

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Successful Clinical Pregnancy and Livebirth was Achieved after Intracytoplasmic Sperm Injection in a Globozoospermic Patient: A Case Report (Globozoospermia를 가진 불임환자에서 미세정자주입술을 이용한 체외수정시술 후 임신과 분만 1례 보고)

  • Kim, Eun-Kyung;Kim, Eun-Ha;Kim, Eun-Ah;Kwon, Hwang;Hong, Jae-Yup;Choi, Dong-Hee
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.1
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    • pp.83-88
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    • 2010
  • Objective: Globozoospermia is observed in <0.1% of male infertile patients. The aim of this study is to report the successful clinical outcome in man with globozoospermia. Design: A case report. Method: A 36-year-old man and 32-year-old partner visited our fertility center with 6 years infertility duration. The semen analysis revealed globozoospermia combined with oligozoospermia which was stained with Diff-Quick method. ICSI (Intracytoplasmic sperm injection) was performed on 12 matured oocytes. Result: 5 injected oocytes were fertilized normally. 5 embryos were transferred on day 3 after oocyte retrieval. The patient became pregnant and delivered a healthy boy at 39 weeks of gestation. Conclusion: In case of globozoospermia, it has a low fertilization rate even though ICSI method is used. The favorable technique is still needed to increase the fertilization rate.

A Study of the Retrovirus-Mediated Transgenic Chicken Production on Chicken Embryos (닭 수정란에서 Retrovirus를 이용한 형질전환 닭 생산 연구)

  • Byun S. J.;Park C.;Kim S. W.;Park J. K.;Chang W. K.;Yang B. S.;Kim T. Y.;Sohn S. H.;Kim S. H.;Jeon I. S.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.225-229
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    • 2005
  • Microinjection of recombinant retrovirus beneath the blastoderm of non-incubated chicken embryo is now the most widespread method for generating transgenic chickens, but transgenesis rates are very low. So to improve this problem, we first introduced retrovirus vector carrying RSV-GFP gene to an one-cell embryo culture system. To investigate whether retrovirus could work on an one-cell chicken embryo, we microinjected the concentrated retrovirus stocks into the germinal disc of one cell or stage-X chicken embryos. Analysis of reporter gene expression on day 4 embryos showed that GFP expression was observed in the only stage-X chicken embryo but was not in the one-cell embryo group. These results suggest that retrovirus system is the most efficient method to generate transgenic chickens in the stage-X embryo.