• KSBB Journal
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• 제26권3호
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• pp.199-205
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• 2011

Using tetrameric ${\beta}$-galactosidase as a model protein, anchoring motives were screened in Bacillus subtilis spore display system. Eleven spore coat proteins were selected considering their expression levels and the location in the spore coat layer. After chromosomal single-copy homologous integration in the amyE site of Bacillus subtilis chromosome, cotE and cotG were chosen as possible spore surface anchoring motives with their higher whole cell ${\beta}$-galactosidase activity. PAGE and Wester blot of extracted fraction of outer layer of purified spore, which express CotE-LacZ or CotG-LacZ fusion verified the existence of exact size of fusion protein and its location in outer coat layer of purified spore. ${\beta}$-galactosidase activity of spore with CotE-LacZ or CotG-LacZ fusion reached its highest value around 16~20 h of culture time in terms of whole cell and purified spore. After intensive spore purification with lysozyme treatment and renografin treatment, spore of BJH135, which expresses CotE-LacZ, retained only 1~2% of its whole cell ${\beta}$-galactosidase activity. Whereas spore of BJH136, which has cotG-lacZ cassette in the chromosome, retained 10~15% of its whole cell ${\beta}$-galactosidase activity, proving minor perturbation of CotG-LacZ, when incorporated in the spore coat layer of Bacillus subtilis compared to CotE-LacZ. Usage of Bacillus subtilis WB700, of which 7 proteases are knocked-out and thereby resulting in 99.7% decrease in protease activity of the host, did not prevent the proteolytic degradation of spore surface expressed CotG-LacZ fusion protein.

• 미생물학회지
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• 제30권3호
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• pp.207-212
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• 1992

합성세제 분해능이 월등한 Pseudomonas putida 와 프탈산에스테르를 분해하는 Arthobacter spp. 의 세포융합을 통해 합성세제와 프탈산에스테르의 분해능을 모두 갖는 융합균주를 개발하였다. Ampicillin-Lysozyme-EDTA 에 의한 각 균주의 원형진체 생성율은 98.4%-99.9% 이었고 원형질체의 재생율은 5-8%이었다. Fusogen으로 40%PEG4000 을 이용하였을 때 효과적으로 융합이 이루어졌으며 융합율은 $1.8{\times}10^{4} $-2.9{\times}10^{4}$ 이었다. 선별된 융합체는 프탈산에스테르와 ABS 를 모두 분해하였으며 ABS 의 분해능은 모균주보다 약 20% 정도 분해능이 증가하였고 DEHP 의 분해능은 모균주와 비슷한수준이었으나 모균주보다 더 빨리 기질을 분해하였다.

• 한국수정란이식학회지
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• 제12권2호
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• pp.171-179
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• 1997

The objectives of the present study were improvements in the efficiency of developmental rates to morula and blastocyst stages to produce a large number of genetically identical nuclear transplant embryos. The oocytes collected from slaughterhouse ovaries were matured for 24 h and then enucleated and cultured to allow cytoplasmic maturation and gain activation competence. And then the donor embryos were treated for 12 h with 10 $\pi$g /ml nocodazole and 7.5 $\pi$g /ml cytochalasin B to synchronize the cell cycle stage at 26 h after the onset of culture. The blastomeres were transferred into the perivitelline space of the enucleated nocytes and blastomeres and oocytes were fused by electrofusion. The cloned embryos were then cultured in various conditions to allow further development. The age of the recipient(30 vs 40 h) had no significant effect on the fusion rates(82.4 vs 82.1%) and the developmental rates to morula /blastocyst(9.8 vs 11.0%). Effect of Nocodazole treatment on the donor cell cyle synchronization to improve the developmental rates of bovine nuclear transplant embryos was significantly higher than control group(21.4 vs 10.1%, p<0.05). Significant differences were in the percentage of fusion rates(72.9,77.1vs 61.9%) in three types of fusion medium(PBS(+), mannitol and sucrose, p<0.01). The developmental rates of bovine nuclear transplant embryos appeared to be highest in mSOF medium under 5% 0$_2$ condition, but no significant differences were found when compared with TCM199-BOEC and mSOF under two different oxygen ratio(5 and 20%).

• Journal of Plant Biology
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• 제27권4호
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• pp.233-239
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• 1984

Reproductives structures of Pachymeniopsis elliptica (Holmes) Yamade (Rhodophyta, Grateloupiaceae) are investigated. In female gametophyte the carpogonial branch and auxiliary cell are produced in separate accessory branch system, the primary ampullar filament originated from mid-cortical layer. After fertilization, auxiliary cell joined with connecting filament becomes a fusion cell by fusing with several neighboring ampullar cells. The fusion cell produces a gonimoblast initial. It divides into gonimoblast cells, which later convert to carposporangia. In male gametophyte superficial cortical cells of vegetative filament produce two spermatangial mother cells which cut off up to three spermatangia respectively. Tetrasporangial initials are formed from the 6th to 12th cells of the cortical layer in tetrasporophyte, and divided cruciately to form tetrasporangium. Some of the sporangia are, however, divided zonately.

• Bulletin of the Korean Chemical Society
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• 제31권11호
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• pp.3411-3414
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• 2010

• Environmental Engineering Research
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• 제19권1호
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• pp.115-115
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• 2014

• Journal of Plant Biology
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• 제15권3호
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• pp.14-18
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• 1972

This paper includes a review on recent development on protoplast culture, regeneraton of plant from protoplast, and fusion of isolated protoplasts, and also describes the possibility of obtaining interspecific hybrid plants through asexual fusion of protoplasts of cells from distantly related plants which are not crossed by the ordinary sexual method.

• 폴리머
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• 제38권6호
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• pp.702-707
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• 2014

각막 내피세포는 각막 가장 안쪽의 단일 세포층이며, 데스메막 위에 놓여있다. 데스메막은 피브로넥틴, 콜라겐, 라미닌, 단백당 등의 포함하는 세포외 기질이라 불리는 다양한 단백질들로 구성되어 있다. 본 연구에서, 조직공학에서 널리 이용되고 있는 락타이드 글리콜라이드 공중합체를 이용하여 투명한 필름을 제작하였으며, 표면에 다양한 부착 분자들(피브로넥틴, 콜라겐 타입 I, IV, 라미닌, FNC 코팅 믹스)을 코팅한 후, 세포 형태 관찰, 세포 증식 및 부착, ZO-1과 $Na^+/K^+-ATPase$의 발현을 확인하였으며, RT-PCR을 통해 각막 내피세포의 인자들을 확인하였다. 실험결과, in vitro 상에서 PLGA 필름은 각막내피세포 전달체로서 역할을 하며 코팅된 세포외 기질들은 각막 내피세포의 거동에 긍정적인 영향을 미침을 확인하였다.

• Journal of Microbiology
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• 제34권1호
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• pp.7-14
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• 1996

Teh baculovirus gp64 glycoprotein is a major component of the envelope of budded virus (BV) and has been shown that it plays an essential role in the infection process, especially virus-cell membrane fusion. We have cloned Autographa californica Nuclear Polyhedrosis Virus (AcNPV) gp64 protein were examined for membrane fusion activity by using a synchtium formation assay under various conditions. The optimal conditions required for inducing membrane fusion are 1) form pH 4.0 to 4.8 2) 15 min exposure of cells to acidic pH 3) at least 1 .mu.g of gp64 cloned plasmid DNA per 3 * 10$^{6}$ cells 4) and an exposure of cells to acidic pH at 72 h post-transfection. In order to investigate the role of hydrophobicity of the gp64 glycoprotein for the membrane fusion, the two leucine residues (amino acid position at 229 and 230) within hydrophobic region I were substituted to alanine by PCR-derived site-directed mutagenisis and the membrane fusion activity of the mutant was anlaysed. The gp64 glycoprotein carrying double alamine substitution mutation showed no significant difference in fusion activity. This result suggested that minor changes in hydrophobicity at the amino acid position 229 and 230 does not affect the acid-induced membrane fusion activity of the gp64 glycoprotein.

• Journal of Plant Biotechnology
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• 제45권3호
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• pp.207-216
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• 2018

Solanum berthaultii is one of the wild diploid Solanum species, which is an excellent resource in potato breeding owing to its resistance to several important pathogens. On the other hand, sexual hybridization between S. berthaultii and S. tuberosum (potato) is limited because of their sexual incompatibility. Therefore, cell fusion can be used to introgress various novel traits from this wild species into the cultivated potatoes. After cell fusion, it is crucial to identify fusion products with the aid of molecular markers. In this study, the chloroplast genome sequence of S. berthaultii obtained by next-generation sequencing technology was described and compared with those of five other Solanum species to develop S. berthaultii specific markers. A total sequence length of the chloroplast genome is 155,533 bp. The structural organization of the chloroplast genome is similar to those of the five other Solanum species. Phylogenic analysis with 25 other Solanaceae species revealed that S. berthaultii is most closely located with S. tuberosum. Additional comparison of the chloroplast genome sequence with those of the five Solanum species revealed 25 SNPs specific to S. berthaultii. Based on these SNPs, six PCR-based markers for differentiating S. berthaultii from other Solanum species were developed. These markers will facilitate the selection of fusion products and accelerate potato breeding using S. berthaultii.