• Journal of Plant Biotechnology
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• 제7권2호
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• pp.113-121
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• 2005

The objective of this study was to identify the major parameters controlling DNA delivery by particle bombardment to immature embryos of Chinese spring wheat (Triticum aestivum L.). Efficiency of DNA (uidA gene) delivery was assessed by transient GUS (${\beta}$-glucuronidase) expression in bombarded tissues. Of the parameters analyzed, acceleration pressure, bombardment distance, chamber vacuum pressure, bombardment times, osmotic conditioning of culture had a remarkable influence on transient gene expression. A bombardment procedure suitable for Chinese spring wheat cultivars was developed which allowed high-efficiency DNA delivery combined with reduced damage to target tissues. The high efficiency made the system practical for wheat genetic transformation research and accelerating wheat breeding programs.

• Clinical and Experimental Reproductive Medicine
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• 제9권1_2호
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• pp.69-77
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• 1982

암모니움 썰페이트 분획분리법. 초미량여과. DEAE - 셀루로오즈 및 젤 여과 크로마토그라피법 등을 사용하여 사람의 정액에서 베타 글룩유로니다아제를 분리하였다. 이 효소는 pH4.4와 온도 50${\sim}$$54^{\circ}C$에서 최고의 활동도를 나타내었다. 파라-나이트로페닐-디-글룩유로나이드를 기질로 사용하였을때 Km치는 0.59mM이었다. 또한 이 효소는 수은이온, 구리이온, 은이온에 의해 활동도가 방해되고 반면에 망간이온 및 코발트 이온에 의해서 효소의 활동도가 증가되었다. 세파댁스 G-200 젤여과법에 의해 측정된 사람의 정액 베타글룩유로니다아제의 분자량은 400,000 이었다. 이렇게 정제된 효소는 수정과정 중에서 카페시태시션의 연구에 도움을 주리라고 본다.

• Journal of Microbiology and Biotechnology
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• 제3권4호
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• pp.285-291
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• 1993

Growth responses of some intestinal bacteria such as bifidobacteria and Clostridium perfringens to the extracts of certain foodstuffs were investigated in vitro. Among edible mountain herbs, the extracts of several chui-na-muls (Aster tataricus, Ligularia fischeri and Aster scaber) had an inhibitory activity against C. perfringens on the agar plate and the water extract of Aster scaber worked selectively on it among intestinal bacteria. The water extract showed growth-promoting effect toward bifidobacteria such as B. adolescentis, B. animalis, B. bifidum, B. infantis and B. thermophilum in the broth culture. When the faecal inoculum was incubated in the culture with the extract, the population of C. perfringens decreased, whereas that of bifidobacteria increased by $10^3$ scale.$\beta$-glucuronidase activity in the culture with the water extract of Aster scaber digested with pepsin and pancreatin was lower than that in the control culture.

• 한국어병학회지
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• 제22권3호
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• pp.305-316
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• 2009

Haemocyte characterization in the surf clam, Mectra veneriformis was carried out based on morphological, cytochemical, and phagocytic characteristics. The haemocytes were classified into two cell types, granulocytes and agranulocytes on the basis of presence of cytoplasmic granules. Granulocytes were then classified again into 2 types, large eosinophilic granulocytes and small eosinophilic granulocytes after staining with May-Grünwald Giemsa. In electron microscopy, both types of granulocytes contained electron-dense and electron-lucent cytoplasmic granules. Agranulocytes (hyalinocytes) were also divided into two cell types, large agranulocytes and small agranulocytes based on their sizes. Both cell types did not have granules in cytoplasm. Granulocyte and agranulocyte were negative for the enzyme activities of alkaline phosphatase, peroxidase, and $\beta$-glucuronidase but positive for phenoloxidase and acid phosphatase activities. Both types of haemocytes have phagocytic activity, with the exception of small agranulocyte, and granulocytes seemed more active in this respect than agranulocytes. This present study is the first study to characterize haemocytes of M. veneriformis.

• Plant Biotechnology Reports
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• 제2권4호
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• pp.267-270
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• 2008

We have successfully used the low-pressure BioWare gene gun, developed for gene transfer in animal cells, for plant tissues. The BioWare device is easy to manipulate. Just 50 psi helium pressure was sufficient to transfer foreign genes into the aleurone layer and embryo of maize without causing tissue damage in the impact area. As shown by expression signals from invasive histochemical ${\beta}-glucuronidase$ (GUS) activity, the foreign reporter gene expressed well in bombarded tissues. This successful GUS-transient expression extends the application of this low-pressure gene gun from animal cells to plant tissues.

• Fisheries and Aquatic Sciences
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• 제9권4호
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• pp.135-139
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• 2006

We optimized the biological and physical parameters for DNA delivery into thalli of the red alga Porphyra yezoensis using a particle bombardment device. The efficiency of transformation was determined using the ${\beta}-glucuronidase$ (GUS) assay. The optimal helium pressure, distance of tungsten particle flight, and ratio of DNA to tungsten particles were $23kgf/cm^2$, 8 cm, and $5{\mu}g/mg$ tungsten, respectively. During bombardment, osmotic treatment with a mixture of 0.6 M mannitol and sorbitol increased the efficiency of GUS transformation. After 2 days, the blue color indicating GUS activity was observed using a histochemical assay.

• KSBB Journal
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• 제6권4호
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• pp.385-388
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• 1991

자란(B. striata)의 미성숙종자로 부터 유도한 embryogenic callus를 현탁배양하고 이와같은 embryogenic cell suspensions로부터 분리한 원형질체에 electroporation 방법을 사용하여 reporter genes이 들어있는 pBI121 plasmid DNA 를 도입하고 그 발현을 확인하였다. 배양된 세포에 있어서 GUS의 활성은 plasmid DNA양의 증가와 함께 높게 나타났으며, 200-300 voltage/1180 uF에서 GUS의 활성 및 원형질체의 생존율(viability)이 가장 좋았다.

• 아시안잔디학회지
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• 제18권4호
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• pp.211-219
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• 2004

Transgenic zoysiagrass (Zoysia japonica cv. Zenith) plants have been obtained by particle bombardment of embryogenic callus with the plasmid pSMABuba, which contains hygromycin resistance (hpt) and bialaphos resistance (bar) genes. Parameters on DNA delivery efficiency of the particle bombardment were partially optimized using transient expression assay of a chimeric $\beta-glucuronidase$(gusA) gene driven by the CaMV 35S promoter. Stably transfarmed zoysiagrass plants were recovered with a selection scheme using hygromycin. Transgenic zoysiagrass plants were confirmed by PCR analysis with specific primer for bar gene. Expression of the transgene in transformed zoysiagrass plants was demonstrated by Reverse transcriptase (RT)-PCR analysis. All the tested transgenic plants showed herbicide BastaR resistance at the field application rate of $0.1\%-0.3\%$.

• Journal of Plant Biology
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• 제37권1호
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• pp.17-25
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• 1994

Two potato (Solanum tuberosum L.) cultivars were transformed by Agrobacterium tumefaciens harboring cauliflower mosaic virus (CaMV) 35S promoter and $\beta$-glucuronidase (GUS) gene. Expression patterns of the CaMV 35S promoter according to tissue types and developmental stages, and genetic stability of GUS gene were investigated in the clonal progenies of transgenic potatoes. Kanamycin-resistant shoot emerged from tuber disc after 4 weeks of culture, and root was induced 6 weeks after culture on the selection medium. Shooting frequency of cvs. Superior and Dejima were 43% and 27%, respectively. Mature transformants and their clonal progenies showed no phenotypical abnormality. GUS activity was expressed primarily at parenchymatous cells of phloem tissue around the vascular cambium in the stem and root, and higher activity was found at the apical meristem of shoot, root and adventious shoot bud. GUS activity was higher at tubers of young explants than at stored tubers. These facts indicate that expression level of the CaMV 35S promoter differed according to tissue types and developmental stages of the organs. The GUS gene was stably inherited to each clonal progeny and normally expressed.

• 농업생명과학연구
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• 제45권6호
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• pp.193-199
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• 2011

An efficient transient expression system has been developed and characterized for the production of foreign genes in seedlings. The seedlings can be easily produced from commercial seeds used for vegetable sprouts. In principal, a chemical abrasive was employed to generate wounds in seedlings prior to vacuum-infiltration with Agrobacterium tumefaciens bearing the target gene. This optimized chemical wounding-assisted agro-infiltration process resulted in up to 15-fold increase in $\beta$-glucuronidase (GUS) enzyme activity. This procedure has been used efficiently to express hepatitis B surface antigen (HBsAg) protein in a transient mode. Therefore, seedlings with proper wounds can be suggested as a convenient tool for the production of useful recombinant proteins.