• Title/Summary/Keyword: Expression vector

Search Result 1,533, Processing Time 0.028 seconds

Retroviral Gene Expression in Spermatogonial Stem Cells during Long-term Culture

  • Jeong, Dong Kee;Griswold, Michael D.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.20 no.7
    • /
    • pp.1015-1022
    • /
    • 2007
  • The spermatogonial stem cell (SSCs) is unique in that it is the only cell in the adult male that can contribute genes to a subsequent generation. Permanent modification of the germ cell line may be realized if stem cells could be cultured, transfected with unique genes, and then transplanted into recipient testes. We developed a culture system that supported long-term viability of SSCs. We used a retrovirus vector (pMSCV including ${\beta}$-galactosidase) to stably transfect spermatogonia following long-term culture using the system developed. Expression of the reporter gene ${\beta}$-galactosidase controlled by the retroviral vector was stable in long-term cultured SSCs. We confirmed the retroviral-mediated ${\beta}$-galactsidase gene could be expressed in germ cells in recipient mice following SSCs transplantation.

Enhanced p53 Gene Transfer to Human Ovarian Cancer Cells using Cationic Nonviral Vector. DOC

  • Choi, Eun-Jeong;Choi, Sung-Hee;Park, Jeong-Sook;Ahn, Woong-Shick;Kim, Chong-Kook
    • Proceedings of the PSK Conference
    • /
    • 2002.10a
    • /
    • pp.427.2-428
    • /
    • 2002
  • Previously we formulated new cationic liposomes, DDC, composed of DOTAP. DOPE, and cholesterol (Chol) in 1 : 0.7 : 0.3 molar ratios, and showed that DDC efficiently deliver the plasmid DNA into ovarian cancer cell lines. Here, wild type p53 DNA was transfected into ovarian cancer cells, using the DOC as a nonviral vector and the expression and activity of p53 gene were evaluated in vitro and in vivo. The complexes of plasmid DNA (pp53-EGFP) and DDC were transfected into OVCAR-3 cells. The gene expression was determined by RT -PCR and western blot analysis. (omitted)

  • PDF

Enhancement of chitinolytic activity of by co-expression of endochitinase and chitobiosidase genes (Endochitinase와 Chitobiosidase 유전자의 동시발현에 의한 키틴분해 활성의 증가)

  • Kim, Jungtae;Choi, Shin-Geon
    • Journal of Industrial Technology
    • /
    • v.30 no.B
    • /
    • pp.69-74
    • /
    • 2010
  • Chitinolytic activity was enhanced by coexpression of endo-chitinase gene (chiA) and chitobiosidase gene (chiB) from Serratia marcescens KFRI314 using constitutive expression vector, pHCEIA, in E. coli. Coexpression vector was constructed by inserting ribosome binding site (RBS) into junction between two chitinase genes. SDS-PAGE analyses showed that two chitinase were constitutively expressed while E. coli clones expressing two chitinases simultaneously increased halo size on colloidal chitin plate. Furthermore, the chitinolytic activities were much enhanced in coexpressed clones when degradation patterns of substrate analogues such as 4-MU-(NAG), $4-MU-(NAG)_2$,$4-MU-(NAG)_3$ were used. Consequently, the combined use of endochitinase and chitobiosidase greatly increased overall chitinolytic activities on recombinant E. coli clones.

  • PDF

Replication and encapsidation of recombinant Turnip yellow mosaic virus RNA

  • Shin, Hyun-Il;Kim, In-Cheol;Cho, Tae-Ju
    • BMB Reports
    • /
    • v.41 no.10
    • /
    • pp.739-744
    • /
    • 2008
  • Turnip yellow mosaic virus (TYMV) is a positive strand RNA virus that infects mainly Cruciferae plants. In this study, the TYMV genome was modified by inserting an extra subgenomic RNA promoter and a multiple cloning site. This modified TYMV was introduced into Nicotiana benthamiana using a Agrobacterium-mediated T-DNA transfer system (agroinfiltration). When a gene encoding $\beta$-glucuronidase or green fluorescent protein was expressed using this modified TYMV as a vector, replication of the recombinant viruses, especially the virus containing $\beta$-glucuronidase gene, was severely inhibited. The suppression of replication was reduced by co-expression of viral silencing suppressor genes, such as tombusviral p19, closteroviral p21 or potyviral HC-Pro. As expected, two subgenomic RNAs were produced from the recombinant TYMV, where the larger one contained the foreign gene. An RNase protection assay revealed that the recombinant subgenomic RNA was encapsidated as efficiently as the genuine subgenomic RNA.

Yeast Cloning Vectors and their Application to the Development of Starch-fermenting Yeast (효모 Cloning Vector와 전분발효 효모의 개발)

  • Kim, Keun
    • Applied Biological Chemistry
    • /
    • v.31 no.3
    • /
    • pp.267-273
    • /
    • 1988
  • Transformed, hybrid strains of the yeast Saccharomyces capable of simultaneous secretion of both glucoamylase and ${\alpha}-amylase$ have been produced. These strains can carry out direct, one-step assimilation of starch with conversion efficiency greater than 93% during a 5 day growth period. One of the transformants converts 92.8% of available starch into reducing sugars in only 2 days. Glucoamylase secretion by these strains results from expression of one or more chromosomal STA genes derived from Saccharomyces diastaticus. The strains were transformed by a plasmid(pMS12) containing mouse salivary ${\alpha}-amylase$ cDNA in an expression vector containing yeast alcohol dehydrogenase promoter and a segment of yeast $2{\mu}$ plasmid. The major starch hydrolysis product produced by crude amylases found in culture broths is glucose, indicating that ${\alpha}-amylase$ and glucoamylase act cooperatively.

  • PDF

Expression of Recombinant Human Bone morphogenetic protein 2 (hBMP2) in Insect cells

  • Kim, Seong-Wan;Kim, Seong-Ryul;Park, Seung Won;Goo, Tae-Won;Choi, Kwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • v.34 no.1
    • /
    • pp.1-5
    • /
    • 2017
  • Bone morphogenetic protein 2 (BMP2) plays an important role in the development of bone and cartilage. It is involved in the hedgehog pathway, TGF beta signaling pathway, and in cytokine-cytokine receptor interaction. It is involved also in cardiac cell differentiation and epithelial to mesenchymal transition. In this study, We expressed human BMP2 (hBMP2) recombinant protein using Baculovirus Expression Vector System (BEVS) in Sf9 insect cells. The hBMP2 cDNA was cloned into baculovirus transfer vector, pBacgus-4x-1 and recombinant baculovirus was screened out through X-gal and GUS-fusions assay. Western blot analysis shown that molecular weight of hBMP2 recombinant protein was about 44.71 kDa.

Construction of Binary Vectors for the Rice Transformation Using a Rice Actin Promoter and Replication Origin of pTi12 Isolated from Agrobacterium tumefaciens KU12 (Agrobactrium tumefaciens KU12로부터 분리한 pli12의 Replication Origin과 벼의 Actin 유전자 프로모터를 이용한 벼의 Binary Vector 제조)

  • Sim, Woong-Seop
    • Journal of Plant Biology
    • /
    • v.38 no.4
    • /
    • pp.365-371
    • /
    • 1995
  • Binary vectors, pBI-ActR1, pBI-ActF1 and pBSH-ActR1, were constructed using pGA642, the replication origin of pTi12 and the rice actin promoter. The sizes of pBI-ActR1, pBI-ActF1 and pBSH-ActR1 were 12.9 kb, 13.2 kb and 11.95 kb, respectively. These vectors containing a rice actin promoter followed by a GUS structural gene could induce stronly the expression of GUS gene in transformed rice cells. Rice explants from 3-4 day old seedlings after germinatin were cocultured with A. tumefaceins harboring pBI-ActR1, pBI-ActF1 or pBSH-ActR1, and then GUS expression in the explants was assayed. Transformation of rice explants by these binary vectors was tissue-specific, such that the meristematic regions of shoot apex, root and hypocotyl were transformed by these binary vectors.

  • PDF

Generalized Vector Control with Reactive Power Control for Brushless Doubly-Fed Induction Machines

  • Duan, Qiwei;Liu, Shi;Schlaberg, H. Inaki;Long, Teng
    • Journal of Power Electronics
    • /
    • v.18 no.3
    • /
    • pp.817-825
    • /
    • 2018
  • In this paper, a current hysteresis control with good decoupling properties for doubly-fed brushless induction machines (BDFIMs) has been proposed based on a generalized vector model. The independent control of the reactive power and speed for BDFIMs has been achieved by controlling the d-axis and the q-axis current of the control windings (CW). The proposed vector control method has been developed for the power winding (PW) flux frame. Experimental verification of a type Y180M-4 BDFIM prototype with 1/4 pole-pairs has been presented. Evidence of its good performance has been shown through experimental results.

Hangul Font Outline Vector Modification Algorithm According to Weather Information (날씨에 따른 한글 폰트 윤곽선 벡터 변형 알고리즘)

  • Park, Dong-Yeon;Jo, Se-Ran;Kim, Nam-Hee;Lim, Soon-Bum
    • Journal of Korea Multimedia Society
    • /
    • v.25 no.9
    • /
    • pp.1328-1337
    • /
    • 2022
  • Recently, research on various font designs has been actively conducted to deliver effective emotional information in a digital environment. In this study, we propose a Hangul font outline vector modification algorithm that effectively conveys sensitivity according to weather information and can be transformed immediately. The algorithm performs a series of transformations: sets outlines according to design pattern templates, calculates the glyph's position to reflect physical rules, splits outline segments into smaller sizes and deforms the outlines. Through this, we could create several vector font designs such as humidity, cloud, wind, and snow. The usability evaluation was close to good, so it can be used in diverse ways if we improve readability and effective design expression.

Regulation of Gene Expression in Higher Plant (고등식물의 유전자 발현의 조절)

  • 심웅섭
    • Proceedings of the Botanical Society of Korea Conference
    • /
    • 1987.07a
    • /
    • pp.241-260
    • /
    • 1987
  • The regulatory mechanisms of gene expression in higher plant were not ascertained in detail because the genome size is very large and complex. However, the above-mentioned study is remarkably progressed in parallel with development of DNA recombinant technology and plant vector system. Some research results connected with the mechanisms could be summarized as follows. 1. Many plant genes including chloroplast genes are cloned. 2. The structures of some regulatory regions of gene expression are determined, and it is confirmed that new regulatory units are made by transposable elements. 3. Plant gene expression is regulated not only at transcriptional level but also at translational level. 4. The factors that regulate plant gene expression could be divided as two categorys. One is endogenous elements including the structural change of chromatin during development stage and tissue differentiation. The other is environmental stimulations such as air, water, heat, salts and light. However, some sufficient research-aid fund is essential in order to study the regulatory mechanisms of gene expression more systematically.

  • PDF