• 한국수산과학회지
• /
• 제49권5호
• /
• pp.628-634
• /
• 2016

We investigated the characteristics and rate of development of allotriploid embryos derived from a cross between female olive flounder Paralichthys olivaceus and male starry flounder Platichthys stellatus. The allotriploidy was induced by cold shocking fertilized eggs three minutes post-fertilization at 3°C for 45 minutes. The average cellular DNA content of the allotriploid embryos was 2.06±0.03 pg/cell, which is equal to the sum of the cellular DNA content of a diploid olive flounder (1.42 pg/cell) and a haploid starry flounder (0.66 pg/haploid cell). The first cleavage, midblastula, gastrula and Kupffer's vesicle appearance stages of the allotriploid eggs began at 1.5, 8, 13 and 26 hours after cold shocking at 18°C, respectively. The developmental rate of allotriploid eggs was equivalent to that of diploid and triploid olive flounder eggs at 10, 14 and 18°C. However, the hatching times of allotriploid eggs, 7 h at 10°C, 5 h at 14°C and 4 h at 18°C, were earlier than those of diploid and triploid olive flounder.

• Journal of Microbiology and Biotechnology
• /
• 제6권6호
• /
• pp.386-390
• /
• 1996

The ${\beta}$-galactosidase gene from Lactococcus lactis subsp. lactis ATCC 7962 was cloned and its enzymatic properties were characterized, with a view to assessing its potential use as a selection marker in the food-grade cloning vector. Chromosomal DNA from L. lactis subsp. lactis 7962 was cleaved with PstI and ligated into pBR322 for transformation into Escherichia coli TGl. Transformants showing ${\beta}$-galactosidase activity possessed the pBR322 plasmid containing a 10 kilobase (kb) PstI fragment and this plasmid was named pCKL11. The cloned ${\beta}$-galactosidase gene came from the chromosomal DNA of L. lactis subsp. lactis 7962 was confirmed by Southern hybridization. A restriction map of pCKL11 was constructed from the cleavage of both pCKL11 and the purified 10kb insert fraqment. The. optimum pH of the ${\beta}$-galactosidase determined with the E. coli harboring the pCKL11 was 7.0. The optimum temperature was $50^{\circ}C$, while the pI of the enzyme was 7.4. These values were the same as those of the enzyme from the parent strain.

• 약학회지
• /
• 제47권5호
• /
• pp.319-324
• /
• 2003

This study examined the inhibitory effect of extracts of Trichosanthes kirilpwii sorted according to the parts on the growth of HL-60 cells. The growth of HL-60 leukemia cells was markedly inhibited by the treatment of the 80％ methanol extract of roots (10 $\mu\textrm{g}$/mι), stems (50$\mu\textrm{g}$/mι), pips (10$\mu\textrm{g}$/mι), and gourds (100 $\mu\textrm{g}$/mι), or the ethylacetate fraction of leaves (100 $\mu\textrm{g}$/mι). when the HL-60 cells were treated with the extracts of T. kirilpwii sorted according to the parts, DNA fragmentation and sub-G1 hypodiploid cells were observed. Moreover, T. kirilpwii extracts increased the level of the expression of the active form of caspase-3 and the activation of caspase-3 was demonstrated by the cleavage of poly(ADP-ribose) polymerase, a vital substrate of effector caspase. The results suggest that the inhibitory effect of extracts of T. kirilpwii sorted according to the parts on the growth of HL-60 cells seems to arise from the induction of apoptosis.

• 한국동물생명공학회지
• /
• 제37권2호
• /
• pp.80-86
• /
• 2022

The ability to determine the sex of bovine embryos before the transfer is advantageous in livestock management, especially in dairy production, where female calves are preferred in milk industry. The milk production of female and male cattle benefits both the dairy and beef industries. Pre-implantation sexing of embryos also helps with embryo transfer success. There are two approaches for sexing bovine embryos in farm animals: invasive and non-invasive. A non-invasive method of embryo sexing retains the embryo's autonomy and, as a result, is less likely to impair the embryo's ability to move and implant successfully. There are lists of non-invasive embryo sexing such as; Detection of H-Y antigens, X-linked enzymes, and sexing based on embryo cleavage and development. Since it protects the embryo's autonomy, the non-invasive procedure is considered to be the safest. Invasive methods affect an embryo's integrity and are likely to damage the embryo's chances of successful transformation. There are different types of invasive methods such as polymerase chain reaction, detection of male chromatin Y chromosome-specific DNA probes, Loop-mediated isothermal amplification (LAMP), cytological karyotyping, and immunofluorescence (FISH). The PCR approach is highly sensitive, precise, and effective as compared to invasive methods of farm animal embryonic sexing. Invasive procedures, such as cytological karyotyping, have high accuracy but are impractical in the field due to embryonic effectiveness concerns. This technology can be applicable especially in the dairy and beef industry by producing female and male animals respectively. Enhancing selection accuracy and decreasing the multiple ovulation embryo transfer costs.

• Journal of Microbiology and Biotechnology
• /
• 제32권12호
• /
• pp.1547-1552
• /
• 2022

β-Amyrin is a pentacyclic triterpene widely distributed in leaves and stems worldwide. The ability of β-amyrin to induce the production of reactive oxygen species (ROS) in microorganisms suggests its potential as an antimicrobial agent. Thus, this study aimed to elucidate the antibacterial mode of action of β-amyrin. We treated Escherichia coli cells with β-amyrin and found that it triggered ROS accumulation. Excessive stress caused by ROS, particularly hydroxyl radicals, induces glutathione (GSH) dysfunction. GSH protects cells from oxidative and osmotic stresses; thus, its dysfunction leads to membrane depolarization. The resultant change in membrane potential leads to the release of apoptotic proteins, such as caspases. The activated caspases-like protein promotes the cleavage of DNA into single strands, which is a hallmark of apoptosis-like death in bacteria. Apoptotic cells usually undergo events such as DNA fragmentation and phosphatidylserine exposure, differentiating them from necrotic cells, and the cells treated with β-amyrin in this study were positive for annexin V and negative for propidium iodide, indicating apoptosis-like death. In conclusion, our findings suggest that the antibacterial mode of action of β-amyrin involves the induction of ROS, which resulted in apoptosis-like death in E. coli.

• 생명과학회지
• /
• 제24권10호
• /
• pp.1078-1084
• /
• 2014

본 연구는 꽈리허리노린재(Acanthocoris sordidus) 추출물이 산화적 스트레스에 의해 유발되는 세포와 DNA의 손상 억제력을 조사하였다. 꽈리허리노린재 추출물의 DPPH 유리 라디칼과 수산화 라디칼 제거능은 양성 대조군에 비해 각각 48.9%, 37.8%로 나타났으며, $Fe^{2+}$-chelating 효과는 80.0%로 조사되었다. 꽈리허리노린재 추출물이 활성산소에 의해 유도되는 세포손상에 미치는 억제 효과를 조사하기 위해 지질과산화의 상대적 수준과 p21 단백질의 발현율을 조사해보면 꽈리허리노린재 추출물은 라디칼 처리군에 비해 지질과산화를 거의 완벽하게 억제하고 있으며, p21 단백질의 발현은 대조군의 92.4%로 회복되는 것으로 조사되었다. 또한 꽈리허리노린재 추출물의 DNA 분절화 억제 활성은 대조군에 비해 53.3%로 나타나 산화적 스트레스에 의해 유발되는 DNA 분절화를 효율적으로 억제하고 있으며, H2AX 단백질의 인산화비는 라디칼 처리군의 39.0%에 해당하는 수준으로 조사되어 꽈리허리노린재 추출물은 히스톤 단백질의 인산화를 매우 효율적으로 억제하는 것으로 확인되었다. 이러한 결과로 보아 꽈리허리노린재 추출물은 활성산소에 대한 항산화 효과뿐만 아니라 세포와 DNA의 손상을 억제하는데 효과적인 것으로 사료된다.

• 대한본초학회지
• /
• 제32권3호
• /
• pp.45-53
• /
• 2017

Objective : Nelumbo nucifera, its rhizome and semen have been used as a traditional medicine which was studied on antioxidant, hepatoprotective effect, anti-obesity and the others. However, Nelumbinis Flos have not studied. We investigated protective effects on oxidative DNA damage and anti-inflammatory effects of Nelumbinis Flos. Methods : The antioxidant activity was conducted by 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, 2, 2'-Azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) radical scavenging assay and reducing power assay. Total phenolic content was analyzed. Also, phenolic compounds were detected by HPLC/UV. The inhibitory effect on oxidative DNA damage was determined using ${\Phi}X-174$ RF I plasmid DNA cleavage assay. The anti-inflammatory effect of Nelumbinis Flos was measured by the amount of nitric-oxide (NO) produced and protein levels of iNOS, and COX-2 in LPS induced RAW 264.7 cells. Results : The results of DPPH and ABTS radical scavenging activity at $200{\mu}g/m{\ell}$ of extraction were $97.02{\pm}0.88%$ and $96.42{\pm}0.25%$. Reducing power (fold of L-ascorbic acid as control) was $100.14{\pm}0.31$ at $200{\mu}g/m{\ell}$. Total phenol content was $8.70{\pm}0.02mg/g$. Chlorogenic acid, catechin and epicatechin were found by HPLC. Nelumbinis Flos has inhibitory effect in dose-manner against oxidative DNA damage. In addition, it showed the anti-inflammatory effect by suppression of NO production as well as protein levels of iNOS, and COX-2. Conclusion : This study suggested that Nelumbinis Flos showed potential antioxidant and suppression activities of various factors were related in NO produced. Therefore, Nelumbinis Flos as natural plant resources that may help reduce inflammation and alleviate DNA damage.

• 미생물학회지
• /
• 제31권2호
• /
• pp.141-147
• /
• 1993

백색 부후균인 plaurotus ostreatus 의 4가지의 균주로부터 각각 10.2 kb 와 7.2 kb (NFFA 2), 두 종류의 10.2 kb (NFFA 4001) 11.2 kb (NFFA 4501), 10.2kb 와 11.2 kb(KFCC 11635) 크기의 미토콘드리아 플라스미드들을 분리해 내었다. NFFA 2 의 변종인 NFFA 2m1 과 NFFA 2m2 에서는 이들 플라스미드가 관찰되지 않았다. 분별 원심분리에 의해 얻은 미토콘드리아에서 핵산을 추출하여 agarose gel 에서 전기영동시키면 플라스미드가 관찰되지 않았으나 proteinase K 를 처리하고 핵산을 추출하여 전기영동한 결과 이들 플라스미드가 관찰되었는에, 이는 플라스미드상에 단백질인 결합되어 있음을 시사한다. Proteinase K 를 처리한 플라스미드 DNA 와 exonuclease 를 반응시킨 결과, 이들 플라스미드들은 5'말단에 단백질이 결합된 성형 이중가닥 DNA의 구조를 가진 것을 확인하였다. 각 플라스미드들의 상호관계를 조사하기 위하여 Southern hybridization 을 수행한 결과 최소 3가지 종류의 플라스미드들로 분류할 수 있었다. 이중 한 그룹 (group I) 은 모든 P. ostreatus 균주들에서 공통적으로 발견되었다. Pleurotus 속의 5가지 다른 종(P. cornucopiae, P. florida, P. pulmonarius, P. sajor-cuja, P. spodoleucus) 의 균주들로부터 미토콘드리아 플라스미드들을 분리하였다. 이들은 한균주당 1-4 개 까지의 플라스미드를 가지며, 플라스미드의 크기는 7.2 kb-14kb 범위에 있었다. P. ostreatus 의 NFFA 2 의 10.2 kb(group I) 플라스미드와 hybridization 을 수행한 결과 P. cornucopiae ASI 2011을 제외한 다른 모든 균주들이 유사한 염기서열의 플라스미드를 갖고 있음을 알았다.

• 한국미생물·생명공학회지
• /
• 제32권1호
• /
• pp.91-95
• /
• 2004

원유로 오염된 토양의 생물학적 복원과정 동안 접종된 Nocardia sp. Hl7-1 균주를 확인하기 위해 165 rDNA sequence에 기초하여 균주에 특이적인 primer를 제작하였다 14균주의 16S rDNA sequence비교를 통해 제작된 4개의 primer set는 Hl7-1 균주를 특이적으로 검출할 수 있었다. 특히 NH169F-NH972R과 NH575F-NH972R의 primer set는 50 fg의 DNA와 $1.2${\times}$10^4$ cfu/g-soil의 균체농도까지 민감하게 검출할 수 있었다. 이 두 primer set는 원유로 오염된 토양의 bioremediation과정 동안 접종된 Hl7-1 균주의 특이적 검출을 가능케 하였으며, 이는 사용된 primer set에 의해 증폭된 PCR산물을 제한효소(EcoRI)로 절단한 결과와 DGGE를 통한 Hl7-1 균주의 확인을 통해 본 연구에서 제작된 primer set의 특이성을 검증하였다.

• 한국미생물·생명공학회지
• /
• 제40권4호
• /
• pp.348-355
• /
• 2012

본 연구에서는 S. cerevisiae와 P. pastoris에서 bovine pancreatic (bp-) DNase I의 과발현과 재조합 DNase I의 특성을 조사하였다. bp-DNase I 유전자는 GAL10 promoter, $MF{\alpha}$, GAL7 terminator 사이에 삽입하여 재조합 plasmid인 pGAL-$MF{\alpha}$-DNaseI (6.4 kb)를 구축하였다. 그리고 bp-DNase I 유전자를 AOX1 promoter, $MF{\alpha}$, AOX1 terminator 에 삽입하여 재조합 plasmid인 pPEXI (8.8 kb)를 구축하였다. 재조합 plasmid인 pGAL-$MF{\alpha}$-DNaseI과 pPEXI를 각각 S. cerevisiae와 P. pastoris 숙주세포에 형질전환시켰다. 형질전환된 효모세포들을 galactose와 methanol 배지에서 $30^{\circ}C$, 48시간 배양하면 bp-DNase I은 대부분이 배양 상등액으로 과발현되었다. P. pastoris 형질전환체는 배양 상등액에서 45.5 unit/mL의 DNase I 활성을 보였으며, 반면에 S. cerevisiae 형질전환체는 37.7 unit/mL의 DNase I 활성을 보였다. 또한 DNA 분해 특성을 조사한 결과, P. pastoris 재조합 DNase I으로 기질 DNA(calf thymus)를 처리하였을 때 1분 이내 DNA가 분해되는 것을 확인할 수 있었으며 이는 상업용 bp-DNase I과 S. cerevisiae 재조합 DNase I으로 처리했을 때보다 빠른 분해 패턴을 보였다.