• KSBB Journal
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• 제5권3호
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• pp.299-306
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• 1990

대장균과 코리네형 세균간의 shutle vector pECCGI과 pECCD2를 제작하고, plasmid pECCGI과 glycine배지에서 다양한 Corynebacterium glutamicum을 사용하여 전기장 충격법에 의한 형질전환에 있어서 여러 조건을 조사한 결과 세포 현탁액 40ul와 DNA 2ul의 혼합액 사용시 저항 600 obms, 전기장의 세기 12.5kv/cm, DNA양 10ng, 세포수 $4.5$\times$10^8$와 세포회수 시기를 1.0이하의 $A^6^0^0$으로 했을때 $10^6$transformants/ug of DNA의 형질전환 효율을 보였다.

• 한국식품영양학회지
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• 제12권3호
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• pp.233-239
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• 1999

To develop the potential use as new host strain for gene cloning the optimal conditions for transform-ation of alkali-tolerant Bacillus sp. were examined. The Bacillus sp. YA-14 was cultured to late logarith-mic growth phase at 37$^{\circ}C$ in modified SPI medium (pH8.0) containing 0.4% MgSO4 0.5mM CaCl2 1 ml of competent cell was mixed with 0.5$\mu$g of plasmid DNA and incubated with shaking at 37$^{\circ}C$ for 40min. The transformation frequency under the optimal condition was 4.53$\times$10-6 CFU/ml/g plasmid DNA. The electrophresis and stably maintained in the new host.

• Mycobiology
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• 제49권2호
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• pp.95-104
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• 2021

Species of the genus Aspergillus have a variety of effects on humans and have been considered industrial cell factories due to their prominent ability for manufacturing several products such as heterologous proteins, secondary metabolites, and organic acids. Scientists are trying to improve fungal strains and re-design metabolic processes through advanced genetic manipulation techniques and gene delivery systems to enhance their industrial efficiency and utility. In this review, we describe the current status of the genetic manipulation techniques and transformation methods for species of the genus Aspergillus. The host strains, selective markers, and experimental materials required for the genetic manipulation and fungal transformation are described in detail. Furthermore, the advantages and disadvantages of these techniques are described.

• BMB Reports
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• 제28권5호
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• pp.414-419
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• 1995

The retroviruses carrying ${\nu}-myc$ and ${\nu}-raf$ oncogenes were infected into fetal thymic organ culture (FTOC) to study the molecular mechanisms involved in T cell development. T cell lymphomas in the different stages of T cell development were obtained from this culture system. Interestingly, a few cell lines obtained from this system have a lack of transfected oncogenes, however these cells have the characteristics of transformed cells. In spite of the discrete phenotype of these transformed cell lines, the same pattern of recombination of endogenous c-raf genes was detected from Southern blot analysis. We suggest in this regard that the translocation event of thymocytes, or abnormal promoter activity, can cause lymphomagenesis by way of c-raf.

• Plant Resources
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• 제4권3호
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• pp.181-188
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• 2001

We tried to introduce two forsythia genes related in lignan biosynthesis, dirigent protein and pinoresinol/lariciresinol (Ph) reductase, into potatoes for accumulation of lignans in transgenic potatoes. We made binary vectors overexpressing dirigent protein gene and P/L reductase gene driven by a CaMV35S promoter and transformed into potatoes via Agrobacterium mediated transformation. And in order to control the metabolic flux of lignan biosynthesis pathway, we tried to inhibit chalcone synthase genes of potatoes by antisense inhibition technique also. We tried to use PCR screening method for selection of transgenic plants of different vectors. We tried to determine and compare lignan contents from different transgenic potato lines.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1985년도 워크샵 및 심포지엄 북한산국립공원의 식생
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• pp.23-33
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• 1985

Plant cell culture is emerging to express bioactive foreign proteins because it has several advantages in that it is safe, economical, genetically stable and eukaryotic expression system comparing with other expression systems. However several limitations such as slow growth rate, low expression level and lack of well established down stream process need to be answered. As a preliminary approach to produce the immunologically interested molecules through the plant cell culture, we tested if granulocyte-macrophage colony stimulating factors (GM-CSFs) from both murine (mGM-CSF) and human (hGM-CSF) are produced as a biologically active form through plant cell culture. The murine and human GM-CSF genes were cloned into the plant expression vector, pBI121, and Ti-plasmid mediated transformation of tobacco leaves was conducted using Agrobacterium tumefaciens harboring both recombinant GM-CSF (rGM-CSF) genes. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Northern blot analysis indicated the expression of the introduced mGM-CSF gene in both transgenic plant and cell suspension cultures. In addition, the biological activities of both murine and human GM-CSF from plant cell culture were confirmed by measuring the proliferation of the GM-CSF dependent FDC-PI and TF-1 cells, respectively.

• 생명과학회지
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• 제14권6호
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• pp.1018-1022
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• 2004

Agrobacterium을 이용한 lisianthus화분의 형질전환을 위하여 적정조건을 수립하였다. 화분의 발아 및 화분관의 발달은 화분발아배지(pollen germination medium; PCM)에 sucrose를 $7-15\%$ 첨가 시키고 pH조건을 5.5-7.0으로 조절하여 $20^{\circ}C-27^{\circ}C$에서 배양할 경우 성공적으로 이루어졌다. 형질전환을 위하여 Agrobacterium 현탁액을 lisianthus화분배양액에 첨가하여 진공침윤을 20 min실시하였으며 형질전환화분은 조직화학적 분석 그리고 발현되는 GUS mRNA를 이용한 RT-PCR 및 Southern hybridization에 의한 DNA산물 분석 등에 의하여 GUS발현을 확인하였다. 이러한 결과를 통하여 화분을 이용한 일시발현기술을 제시하게 되었다.

• Journal of Plant Biotechnology
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• 제39권1호
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• pp.13-22
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• 2012

Plant regeneration has been optimized increasingly by organogenesis and somatic embryogenesis using a range of explants with tissue culture improvements focusing on factors, such as the age of the explant, genotype, media supplements and $Agrobacterium$ co-cultivation. The production of haploids and doubled haploids using microspores has accelerated the production of homozygous lines in Brassicaceae crop plants. Somatic cell fusion has facilitated the development of interspecific and intergeneric hybrids in sexually incompatible species of $Brassica$. Crop improvement using somaclonal variation has also been achieved. Transformation technologies are being exploited routinely to elucidate the gene function and contribute to the development of novel enhanced crops. The $Agrobacterium$-mediated transformation is the most widely used approach for the introduction of transgenes into Brassicaceae, and $in$ $vitro$ regeneration is a key factor in developing an efficient transformation method in plants. Although many other Brassicaceae are used as model species for improving plant regeneration and transformation systems, this paper focuses on the recent technologies used to regenerate the most important Brassicaceae crop plants.

• 한국미생물·생명공학회지
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• 제23권2호
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• pp.156-163
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• 1995

Bacillus sphaericus 1593 is pathogenic to the larvae of a number of mosquito species that are known as important vectors for the transmission of certain human and animal diseases. As a preliminary experiment for developing a multfunctional B. sphaericus 1593 as a potent antagonist, we investigated the conditions for the protoplast transformation system of B. sphaericus 1593 using the plasmid pGB215-110$\Delta$B. The protoplast of B. sphaericus 1593 were obtained most efficiency by treating the cells with 500 $\mu$g/ml of lysozyme in the SMM buffer containing 0.5 M sucrose at pH 8.0 and 40$\circ$C for 60 minutes. The cell wall was regenerated on the plate containing 1.2% agar and 0.8 M mannitol. Under the best condition for protoplast formation and regeneration established in the work the highest frequency of transformation was achieved with the 40% PEG (M.W 4,000) treatment for 15 minutes of incubation at 4$\circ$C, and subsequently for 120 minutes incubation at 30$\circ$C for phenotypic expression. The highest transformation efficiency were observed at 1.0 $\mu$g/ml of the final concentration of the plasmid DNA and the plasmids were found to be fairly stable since about 70% of the plasmids were maintained after 8 successive daily transfers onto the fresh medium.

• 미생물학회지
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• 제42권1호
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• pp.59-61
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• 2006

먹물버섯의 하나인 Coprinellus congregatus를 대상으로 유전자의 도입을 위한 형질전환실험에서 원형질체를 생성하지 않고 분열자 (oidium)를 사용하는 방법을 확립하였다. 분열자는 20일 이상된 CKMM 한천배지에서 생성되며, 이를 밤샘배양으로 발아를 촉진시킨 상태에서 전기천공방법으로 항생물질 basta에 대한 형질전환을 수행한 결과 10-20 형질전환체/${\mu}g$ DNA의 수율로 형질전환체를 확보할 수 있었다. 이 형질전환체들은 도입된 벡터가 염색체 상에 삽입되어 유전적으로 안정된 상태를 유지하였다.