• 한국수정란이식학회:학술대회논문집
• /
• 한국수정란이식학회 2002년도 국제심포지엄
• /
• pp.101-101
• /
• 2002

GH-transgenic coho salmon (Oncorhynchus kitsutch) juveniles in tGH＊T$_3$and tGH*PTU were fed with the diets containing 1 ug/g fish of 3,5,3'-triiodo-L-thyronine (T$_3$) and 30 ug/g fish of 6-n-propyl-2- thiouracil (PTU), respectively, to assess the effect of these drugs on the change of physiological activity, growth and survival rate in comparison with normal transgenic (tGH＊C) and nontransgenic coho salmon (Wild) for 90 days. Although the daily food intakes of all transgenic (tGH)-groups were higher than Wild, the amount was reduced by exogenous PTU supply. The fred efficiencies of tGH-groups were lower than Wild, but the efficiency was reduced both by T$_3$and PTU. The survival rate of tGH-group was significantly higher than that of Wild, but there was no significant difference among tGH-groups. Although the growth of tGH-coho salmon was faster than Wild. the growth rate of transgenic salmon was increased by exogenous T$_3$, but was reduced by PTU Plasma TT$_4$levels of tGH-groups was approximately 2-fold higher relative to Wild, but there were no difference of plasma TT$_4$levels among tGH-groups. plasma TT$_3$level or tGH-coho salmon was increased by exogenous T$_3$administration, but was reduced by exogenous PTU. In addition, although plasma GH levels of all tGH-groups were higher than that of Wild, the GH level in plasma of transgenic coho salmon was increased by exogenous T$_3$and reduced by exogenous PTU. In the meantime, the transgenic fishes also displayed head, jaw and opercular abnormalities typical of the offsets of this gene construct in coho salmon, indicating that some imbalance in growth processes has been induced. However, the abnormalities of transgenic coho salmon was reduced following exogenous PTU administration.

• Fisheries and Aquatic Sciences
• /
• 제25권1호
• /
• pp.1-11
• /
• 2022

Catfish is one of the most important freshwater fish farming commodities in Indonesia. Higher catfish production can be achieved by cultivating transgenic catfish carrying the growth hormone (GH) gene of African catfish (Clarias gariepinus GH, CgGH). This research focuses on analysis of the presence of the CgGH gene in transgenic G₁, G₂, and G₃ mutiara catfish broodstock, as an indication of stable CgGH inheritance. CgGH gene was isolated using the RNeasy mini kit and RT-PCR. RT-PCR revealed amplicons measuring approximately 600 bp in transgenic G₀, G₁, G₂, and G₃ mutiara catfish. The CgGH consensus sequence similarities ranged from 93.76% to 97.06%, with four functional domain sites (somatotropin-1, somatotropin-2, four α-helix, N-glycosylation, four cysteine residues) of fish GH proteins. The functional domains of fish GH proteins are conserved in G₁, G₂, and G₃ and indicate stable exogenous GH inheritance to produce transgenic catfish strains in each generation.

• 한국지구과학회지
• /
• 제29권3호
• /
• pp.255-262
• /
• 2008

고흥지역 일대에 분포하는 백악기 화강암을 대상으로 석영내에 발달해 있는 아문미세균열을 측정하여 연구지역의 고응력장을 분석하였다. 5개의 시료(GH-1, GH-3, GH-4, GH-5, GH-8)에서 아문미세균열들의 방향을 측정하여 분석한 결과, GH-1, GH-3, GH-4에서 최대수평주응력은 $N60^{\circ}W$와 $N70^{\circ}E$, $N20^{\circ}W,\;N50^{\circ}W$의 방향성이 가장 우세하였으며, N-S와 $N30^{\circ}E$의 방향성도 미약하게 나타났다. GH-5와 GH-8시료에서는 최대수평주응력의 방향 $N40^{\circ}E$와 $N10^{\circ}E$가 가장 우세하였고, $N40^{\circ}W$의 방향성도 미약하게 나타났다 전체적인 최대수평주응력의 방향은 $N60^{\circ}W$가 가장 우세하였으며, $N20^{\circ}W$, $N20^{\circ}E$, $N70^{\circ}E$등의 방향성도 미약하게 나타났다. 이 연구의 결과와 기존의 고등력장에 관한 연구결과를 종합해 볼 때, 백악기 말에서 신생대 3기 초 사이 연구지역에 작용한 최대수평주응력의 방향은 WNW에서 NE로 변화했을 것으로 판단되며, 그 원인으로는 그 당시 동북아시아 일대에 일어났던 복잡한 지구조운동에 기인한 것으로 사료된다.

• 생명과학회지
• /
• 제31권3호
• /
• pp.356-365
• /
• 2021

최근, 본 연구진은 담수 환경 유래 한천 분해 세균인 Cellvibrio sp KY-GH-1 (KCTC13629BP)의 전체 유전체 염기 서열을 분석하여 아가로오스를 L-AHG 및 D-갈락토오스로 가수분해시키는 아가레이즈들을 암호화하는 유전 정보를 탐색하였다. 그 결과, KY-GH-1 균주는 유전체 상의 약 77 kb 길이의 아가레이즈 유전자 클러스터 내에 9개의 β-아가레이즈 유전자들과 2개의 α-네오아가로비오스 가수분해효소(α-NABH) 유전자들을 지닌 것으로 나타났다. 이러한 유전자 정보를 바탕으로 KY-GH-1 균주가 한천을 탄소원으로 자화하기 위해 단량체인 L-AHG와 D-갈락토오스로 분해시키는 공정은, 엔도형 GH16 β-아가레이즈, 엔도형 GH86 β-아가레이즈 등에 의해 개시되어 NA4, NA6, NA8 등을 생성시킨 후, 이들에 대해 엑소형 GH50 β-아가레이즈가 추가로 작용하여 NA2를 생성시키고, 이어서 GH117 α-NABH가 작용하여 생성된 NA2를 단량체 L-AHG와 D-갈락토오스로 분해함으로써 종결되는 것으로 예측되었다. 대장균 발현 시스템과 PET-30a 벡터를 함께 사용하여, KY-GH-1 균주 유래의 GH50 패밀리 β-아가레이즈 유전자들(GH50A, GH50B, GH50C)과 GH117 패밀리 α-NABH 유전자들(GH117A α-NABH, GH117B α- NABH)을 발현시켜 His-태그 재조합 효소단백질들로 확보하여, 이들 효소단백질을 이용하여 효소 활성을 비교 분석한 결과, 재조합 GH50A β-아가레이즈가 세 개의 GH50A 패밀리 β-아가레이즈 동위효소들 중에서 가장 높은 엑소형 β-아가레이즈 활성을 나타내며, 또한 재조합 GH117A α-NABH가 NA2를 L-AHG와 D-갈락토오스로 강력하게 가수분해할 수 있으나 재조합 GH117B α-NABH는 NA2 가수분해 활성이 없음을 확인하였다. 연이어 GH50A β-아가레이즈 및 GH117A α-NABH의 효소 특성을 추가로 조사하였다. 아울러 이들 각 효소가 나타내는, 아가로오스를 분해하여 NA2를 생성시키는 효율성과 NA2를 분해하여 L-AHG 및 D-갈락토오스를 생성시키는 효율성을 평가하였다. 본 총설에서는, L-AHG 및 D-갈락토오스의 양산을 위한 아가로오스의 효소적 가수분해에 성공적으로 활용될 수 있을 것으로 기대되는, 담수 유래 한천 분해 세균 Cellvibrio sp. KY-GH-1 유래의 재조합 GH50A β-아가레이즈 및 GH117A α-NABH의 장점들에 대해 기술한다.

• 생명과학회지
• /
• 제18권11호
• /
• pp.1617-1624
• /
• 2008

키틴과 그 탈아세틸화된 형태인 키토산은 지구 상에 가장 풍부하게 존재하는 바이오매스의 하나이다. 키틴과 키토산은 항균활성, 면역증강, 중금속 흡착 등 다양한 생리활성을 보이고 있으며 식품, 의약품, 환경산업 등에서 다양하게 응용되고 있다. 이러한 키틴/키토산을 가수분해하는 효소들과 그 3차구조, 유전자들이 세균, 고세균, 진핵생물등 모든 생물종에서 보고되어 왔다. 탄수화물을 가수분해하는 효소들은 그 아미노산 서열에 따라 CAZy (Carbohydrate Active Enzymes) 데이터베이스에 분류되었는데 흥미롭게도 최근까지 키틴가수분해효소와 키토산가수분해효소들은 14개의 glycosyl hydrolase (GH) family들로 분류되어 있다(GH2, GH5, GH7, GH8, GH18, GH19, GH20, GH46, GH48, GH73, GH75, GH80, GH84, GH85). 본 총설에서는 새로운 유전자원를 찾기위한 한 방편으로서 최근에 새롭게 분류된 glycosyl hydrolase family의 분류법에 따라 각각의 GH family에 속하는 키틴/키토산가수분해효소의 종류 및 구조, 그리고 그 효소적 특징에 대하여 논하고자 한다.

• KSBB Journal
• /
• 제10권3호
• /
• pp.271-282
• /
• 1995

met-hGH로부터 고정화 ApM에 의해 전환된 천 연형 hGH를 등전위점, 소수성, 전하량의 차이에 따라 3회의 일련의 크로마토그래피를 수행하여 정제하고, 정제된 hGH의 특성을 조사하였다. 정제한 hGH의 비활성은 효소면역측정법으로 분석하였을 때 2 2.75IUjmg이었고, hGH 회수율은 14.1%이었다. 정제한 재조합 hGH는 아미노산 조성, 아미노 말단의 15개의 아미노산 서열 분석, 카르복시 말단의 아미노산 분석, 트럽신 가수분해 랩타이드들의 HPLC 분석등을 통해 아미노말단 에치오닌이 제거된 천연 형 hGH임을 알 수 있었고, SDS-PAGE, native­P PAGE, IEF, RP-HPLC, HSGF 등의 분석 결과 정제한 hGH는 상용품보다 순도가 높다는 것을 알 수 있었으며, 뇌하수체 제거 쥐의 체중 증가와 경골성 장폭의 증가를 통한 활성 측정 결과 정제한 hGH는 상용 hGH와 같은 정도의 활성을 냐타내었으며 우혈청 알부민을 주사한 대조군에 비해 뚜렷한 성장 효 과를 나타냈다. 따라서 본 연구에셔 정제한 hGH는 천연형이며, 높은 순도와 우수한 생리활성을 나타내는 것을 알 수 있었다.

• Animal Bioscience
• /
• 제34권5호
• /
• pp.867-879
• /
• 2021

Objective: Fibronectin 3 (FN3) and immunoglobulin like modules (Ig) are usually collocated beside modular cellulase catalytic domains. However, very few researches have investigated the role of these modules. In a previous study, we have sequenced and analyzed bacterial metagenomic DNA in Vietnamese goats' rumen and found that cellulase-producing bacteria and cellulase families were dominant. In this study, the properties of modular cellulases and the role of a FN3 in unique endoglucanase belonging to glycosyl hydorlase (GH) family 5 were determined. Methods: Based on Pfam analysis, the cellulases sequences containing FN3, Ig modules were extracted from 297 complete open reading frames (ORFs). The alkaline, thermostability, tertiary structure of deduced enzymes were predicted by AcalPred, TBI software, Phyre2 and Swiss models. Then, whole and truncated forms of a selected gene were expressed in Escherichia coli and purified by His-tag affinity column for assessment of FN3 ability to enhance enzyme activity, solubility and conformation. Results: From 297 complete ORFs coding for cellulases, 148 sequences containing FN3, Ig were identified. Mostly FN3 appeared in 90.9% beta-glucosidases belonging to glycosyl hydrolase family 3 (GH3) and situated downstream of catalytic domains. The Ig was found upstream of 100% endoglucanase GH9. Rarely FN3 was seen to be situated downstream of X domain and upstream of catalytic domain endoglucanase GH5. Whole enzyme (called XFN3GH5 based on modular structure) and truncate forms FN3, XFN3, FN3GH5, GH5 were cloned in pET22b (+) and pET22SUMO to be expressed in single and fusion forms with a small ubiquitin-related modifier partner (S). The FN3, SFN3 increased GH5 solubility in FN3GH5, SFN3GH5. The SFN3 partly served for GH5 conformation in SFN3GH5, increased modules interaction and enzyme-soluble substrate affinity to enhance SXFN3GH5, SFN3GH5 activities in mixtures. Both SFN3 and SXFN3 did not anchor enzyme on filter paper but exfoliate and separate cellulose chains on filter paper for enzyme hydrolysis. Conclusion: Based on these findings, the presence of FN3 module in certain cellulases was confirmed and it assisted for enzyme conformation and activity in both soluble and insoluble substrate.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2000년도 춘계학술발표대회
• /
• pp.541-545
• /
• 2000

For the sustained release formulation of recombinant human growth hormone (rhGH), dissociable rhGH aggregates were microencapsulated within poly(D,L-lactic-co-glycolic acid) [PLGA] microparticles. rhGH aggregates with 2 - 3 m Particle diameter were first produced by adding a small volume of aqueous rhGH solution into a partially water miscible organic solvent phase(ethyl acetate) containing PLGA. These rhGH aggregates were then microencapsulated within PLGA polymer phase by extracting ethyl acetate into an aqueous phase pre-saturated with ethyl acetate. The resultant microparticles were 2 - 3 m in diameter similar to the size of rhGH aggregates, suggesting that PLGA polymer was coated around the protein aggregates. Release profiles of rhGH from these microparticles were greatly affected by changing the volume of the incubation medium. The release rhGH species consisted of mostly monomeric form with having a correct conformation. This study reveals that sustained rhGH release could be achieved by microencapsulating reversibly dissociable protein aggregates within biodegradable polymers.

• Asian-Australasian Journal of Animal Sciences
• /
• 제16권8호
• /
• pp.1193-1198
• /
• 2003

An increase in frequency of administration of exogenous growth hormone (GH) or GH-releasing hormone was reported to be a model to increase blood circulating insulin-like growth factor-1 (IGF-1) and to improve growth performance in animals. We have investigated the effect of twice daily administration of GH-releasing peptide-2 (GHRP-2) on growth performance, GH responsiveness and plasma insulin-like growth factor IGF-1 in swine. We administered to eight swine, 3 control and 5 treatment, a twice daily s.c. injections of GHRP-2 ($30{\mu}g/kg\;BW$) for a period of 10 days. Every day blood samples immediately taken before injections of GHRP-2 or saline, at 08:00 h and 16:00 h, were measured for IGF-1 concentrations. Blood samples for GH assay were collected every 20 min on days 1, 6 and 10, from 1 hour before and 3 h after GHRP-2 or saline injections at 08:00 h. GH peak concentrations and GH area under curve (GH AUC) on day 1, 6 and 10 in treatment group of swine were higher than those in control swine (p<0.05). Twice daily administration of GHRP-2 caused a significantly attenuation (p<0.05) of GH peak concentrations ($80.25{\pm}13.87$, $39.73{\pm}5.72$ and $27.57{\pm}6.06ng/ml$ for day 1, 6 and 10, respectively) and GH AUCs ($3,536.15{\pm}738.35$, $1,310.31{\pm}203.55$ and $934.37{\pm}208.99ng/ml$ for day 1, 6 and 10, respectively). However, there was no significant difference in GH peak concentration and GH AUC between day 6 and 10. Plasma IGF-1 concentration levels were higher in treatment than control group of swine (p<0.05) after 3 days of the treatment, and the levels reached a plateau from day 3 to 10 of experiment. Growth performance did not alter by GHRP-2 administration, even though a numerical increase of body weight gain and feed efficiency was observed. These results indicate that twice daily administration of GHRP-2 for 10 days in swine did not significantly influence on growth performance, caused an overall attenuation of GH response, and that elevation of plasma GH concentrations caused by GHRP-2 administration increased plasma IGF-1 concentrations, even though an attenuation of GH response was observed.

• Fisheries and Aquatic Sciences
• /
• 제15권1호
• /
• pp.43-47
• /
• 2012

Growth hormone (GH) transgenesis in fish has the potential to improve aquaculture efficiency and capacity. However, many fast-growing transgenic fish have experienced side effects caused by excess GH expression. To overcome this unwanted issue associated with several GH transgenic mud loach Misgurnus mizolepis lines carrying GH construct driven by a strong ${\beta}$-actin regulator ($pml{\beta}$-actGH), we performed an alternative version of GH autotransgenesis using a weaker but more stable regulator, the mud loach lectin promoter. GH transgenesis with a pmlectGH construct consisting of the mud loach GH gene driven by the 2.3-kb lectin promoter exhibited significant growth stimulation. However, the extent of the growth acceleration in pmlectGH transgenics (six times maximum when assessed 2 months post hatching) was much less than that in transgenic individuals carrying the $pml{\beta}$-actGH construct. Additionally, the extraordinary gigantism that was common in $pml{\beta}$-actGH-transgenic mud loaches was diminished in transgenic loaches harboring the pmlectGH construct. Transgenic founders (pmlectGH) successfully transmitted their transgene into the next generation with up to 41% frequency. Growth stimulation also persisted in the transgenic F1 strains, with a seven-fold increase in maximum body weight at 6 months of age.