• 생명과학회지
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• 제6권3호
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• pp.185-192
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• 1996

박테리오파지 T7 gene 2.5 단백질은 single-stranded DNA 결합 단백질로 박태리오파지 T7의 DNA복제, 재조합, 및 수선에 필수적으로 요구된다. Gene 2.5 protein은 T7의 DNA 합성과 성장에 필수적인 단백질이다. Gene 2.5 Protein이 중요시 되는 이유는 이 단백질이 T7의 다른 복제 필수단백질인 T7의 다른 복제 필수단백질인 T7 DNA polymerase 와 gene 4 protein(helicase/primase)와 서로 상호작용할 것으로 제안되었기 때문이다. (Kim and Richardson, J. Biol. Chem., 1992;1994). 이 단백질의 단백질 상호작용을 가능하게 하는 domain은 carboxyl-terminal domain일 것으로 여러 실험에서 대두되었기에, 이 domain의 특성을 파악하기 위해 야생형과 변이체 gene 2.5 단백질들을 각각 GST에 융합한후 fusion 단백질을 정제하였다. 정제된 이 융합 단백질들의 carboxyl-terminal domain이 T7 복제 단백질들과 상호작용을 조사하는지를 조사하기 위해 affinity chromatography로 이용하였다. 실험 결과, 아생형 GST-gene 2.5 융합단잭질(GST-2.5 (WT))는 T7 DNA polymerase 와 상호작용을 하였지만. 변이형 융합단백질(GST-2.5$\Delta$21C)는 interaction을 하지 못했다. 이 결과는 carbohyl-terminal domain이 단백질-단백질 상호작용을 하는데 직접적으로 관여하는 것을 증명하였다. 또한,GST2.5(WT)는 gene 4 protein(helicase/primase)와 직접 상호작용을 하나. GST2.5$\Delta$21C는 상호작용을 하지 못하는 것으로 나타났다. 따라서 gene 4 proteins와의 상호작용에도 gene 2.5 protein의 carboxyl-terminal domain이 직접 관여 한다는 것이 증명되었다. 이상의 결과에서 gene 2.5 protein은 박테리오파지 T7 의 유전자 목제 시 단백질-단백질 상호작용에 관혀아며, 특히 gene 2.5 protein의 carboxyl-terminal domain이 이러한 상호작용에 직접적으로 관여하는 domain이라는 것을 알 수가 있었다.

• 한국축산식품학회지
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• 제39권3호
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• pp.503-509
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• 2019

The aim of this study was to compare the effects of number of washes and salt addition on the meat quality in protein recovered from Alaska Pollock compared with pork leg. Various properties of protein recovered from Alaska Pollock (C, washed twice, no salt) and pork leg (T1, washed twice, no salt; T2, two washes, salt added; T3, washed four times, no salt; and T4, washed four times, salt added) were assessed in this study. Pork leg samples exhibited better color (more whiteness, less yellowness) than Alaska Pollock samples. In pork leg samples, four washes (T3, T4) during processing yielded whiter, less yellow protein than two washes (T1, T2). Overall, the textural property measures were higher in pork leg samples (T2, T3, and T4) than in other samples. Breaking force, jelly strength, and folding resistance were significantly higher in salt-treated pork leg samples (T2, T4) than in the other samples. Our findings demonstrate that protein recovered from pork leg has better color parameters, and physical strength compared with Alaska Pollock-derived protein. A higher number of wash steps and treatment with salt during processing were furthermore found to yield better color, and physical strength in the protein samples.

• Asian-Australasian Journal of Animal Sciences
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• 제29권11호
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• pp.1616-1624
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• 2016

The aim of present experiment was to investigate the effect of protein reduction in commercial broiler chicken rations with incorporation of de-oiled rice bran (DORB) and supplementation of limiting amino acids (valine, isoleucine, and/or tryptophan) with ration formulation either on total amino acid (TAA) or standardized ileal digestible amino acids (SIDAA). The experimental design consisted of $T_1$, TAA control; $T_2$ and $T_3$, 0.75% and 1.5% protein reduction by 3% and 6% DORB incorporation, respectively by replacing soybean meal with supplemental limiting amino acids to meet TAA requirement; $T_4$, SIDAA control, $T_5$ and $T_6$, 0.75% and 1.5% protein reduction by DORB incorporation (3% and 6%) with supplemental limiting amino acids on SIDAA basis. A total of 360 dold fast growing broiler chicks (Vencobb-400) were divided into 36 homogenous groups of ten chicks each, and six dietary treatments described were allocated randomly with six replications. During 42 days trial, the feed intake was significantly (p<0.05) reduced by TAA factor compared to SIDAA factor and protein factor significantly (p<0.05) reduced the feed intake at 1.5% reduction compared to normal protein group. This was observed only during pre-starter phase but not thereafter. The cumulative body weight gain (BWG) was significantly (p<0.05) reduced in TAA formulations with protein step-down of 1.5% ($T_3$, 1,993 g) compared to control ($T_1$, 2,067 g), while under SIDAA formulations, BWG was not affected with protein reduction of 1.5% ($T_6$, 2,076 g) compared to $T_4$ (2,129 g). The feed conversion ratio (FCR) was significantly (p<0.05) reduced in both TAA and SIDAA formulations with 1.5% protein step-down ($T_3$, 1.741; $T_6$, 1.704) compared to respective controls ($T_1$, 1.696; $T_4$, 1.663). The SIDAA formulation revealed significantly (p<0.05) higher BWG (2,095 g) and better FCR (1.684) compared to TAA formulation (2,028 g; 1.721). Intake of crude protein and all limiting amino acids (SID basis) was higher in SIDAA group than TAA group with resultant higher nitrogen retention (4.438 vs 4.027 g/bird/d). The nitrogen excretion was minimized with 1.5% protein reduction (1.608 g/bird) compared to normal protein group (1.794 g/bird). The serum uric acid concentration was significantly reduced in $T_3$ (9.45 mg/dL) as compared to $T_4$ (10.75 mg/dL). All carcass parameters were significantly (p<0.05) higher in SIDAA formulation over TAA formulation and 1.5% protein reduction significantly reduced carcass, breast and thigh yields. In conclusion, the dietary protein can be reduced by 0.75% with TAA formulation and 1.5% with SIDAA formulation through DORB incorporation and supplementation of limiting amino acids and among formulations, SIDAA formulation was better than TAA formulation.

• 한국수산과학회지
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• 제28권2호
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• pp.209-218
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• 1995

본 연구에서는 유전자 복제기작을 생화학적, 분자생물학적 방법을 사용하여 bacteriphage T7을 대상으로 연구하였다. Bacteriophage T7의 유전자 복제, 재조합, 수선시 필수 단백질로 작용하는 gene 2.5 단백질의 생체내 기능에 대한 유전학적 연구와 단백질을 분리 정제하여 복제 단백질들과의 상호작용에 대한 연구를 수행하였다. 연구결과 gene 2.5 단백질은 DNA복제시 필수 구성단백질로 작용하며, 복제과정에서 유전자 복제에 관여하는 핵심 단백질들인 DNA polymerase, helicase/primase와 직접 단백질-단백질 상호 협동 작용을 하는 r것을 증명하였다. 특히 gene 2.5 단백질의 C-terminal domain이 절편된 변이체의 경우 복제 단백질들과 상호작용이 결여되었다. 따라서 C-terminal domain이 gene 2.5 단백질의 기능에 필수적으로 관여함을 입증하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제12권7호
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• pp.1075-1079
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• 1999

Adult German cross $(German{\times}British{\times}Russian)$ angora rabbits (one year age), 32 in number were divided randomly into four groups $(T_1-T_4)$ with equal sex ratio and fed diets containing $T_1$ groundnut cake (GNC); $T_3$, soyaflakes (SF); $T_4$, sunflower cake (SFC) and $T_2$, a mixture of all the three cakes along with green forage as roughage for a period of 9 months. Nine per cent protein was added from each protein source. Fibre level was maintained by adjusting the level of rice phak in the diets. The diets were iso-nitrogenous and contained similar level of fibre. DMI through roughage was not affected due to source of protein in the diet, however, DMI through concentrate was higher $(p{\leq}0.05)$ with SFC diet, which resulted in higher total feed intake in the group $(T_4)$. Body weights increased up to second shearing, thereafter it decreased due to summer depression. Diet containing soyaflakes sustained higher wool yield whereas, it was lowest $(p{\leq}0.05)$ on SFC diet. Wool attributes (staple length, medullation, fibre diameter) were not affected due to source of protein in the diet. Digestibility of fibre and its fractions (ADF, cellulose, hemicellulose) decreased $(p{\leq}0.05)$ with incorporation of SFC in the diets. Balance of calcium was lowest whereas, that of nitrogen was highest with SFC diet $(T_4)$. Biological value of N and net protein utilization was better when different protein sources were mixed together $(T_2)$. Protein quality of soyaflakes proved better for wool production followed by groundnut cake and mixture of three protein sources. Sunflower cake alone or in combination decreased wool production which may be checked by supplementation of amino acids and energy.

• BMB Reports
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• 제37권2호
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• pp.260-267
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• 2004

Transducin (T), the heterotrimeric guanine nucleotide binding protein in rod outer segments, serves as an intermediary between the receptor protein, rhodopsin, and the effector protein, cGMP phosphodiesterase. Labeling of T with dansyl chloride (DnsCl) inhibited its light-dependent guanine nucleotide binding activity. Conversely, DnsCl had no effect on the functionality of rhodopsin. Approximately 2-3 mol of DnsCl were incorporated per mole of T. Since fluoroaluminate was capable of activating DnsCl-modified T, this lysine-specific labeling compound did not affect the guanine nucleotide-binding pocket of T. However, the labeling of T with DnsCl hindered its binding to photoexcited rhodopsin, as shown by sedimentation experiments. Additionally, rhodopsin completely protected against the DnsCl inactivation of T. These results demonstrated the existence of functional lysines on T that are located in the proximity of the interaction site with the photoreceptor protein.

• IMMUNE NETWORK
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• 제11권5호
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• pp.268-280
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• 2011

Background: Dengue virus, which belongs to the Flavivirus genus of the Flaviviridae family, causes fatal dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) with infection risk of 2.5 billion people worldwide. However, approved vaccines are still not available. Here, we explored the immune responses induced by alternating prime-boost vaccination using DNA vaccine, adenovirus, and vaccinia virus expressing E protein of dengue virus type 2 (DenV2). Methods: Following immunization with DNA vaccine (pDE), adenovirus (rAd-E), and/or vaccinia virus (VV-E) expressing E protein, E protein-specific IgG and its isotypes were determined by conventional ELISA. Intracellular CD154 and cytokine staining was used for enumerating CD4+ T cells specific for E protein. E protein-specific CD8+ T cell responses were evaluated by in vivo CTL killing activity and intracellular IFN-${\gamma}$ staining. Results: Among three constructs, VV-E induced the most potent IgG responses, Th1-type cytokine production by stimulated CD4+ T cells, and the CD8+ T cell response. Furthermore, when the three constructs were used for alternating prime-boost vaccination, the results revealed a different pattern of CD4+ and CD8+ T cell responses. i) Priming with VV-E induced higher E-specific IgG level but it was decreased rapidly. ii) Strong CD8+ T cell responses specific for E protein were induced when VV-E was used for the priming step, and such CD8+ T cell responses were significantly boosted with pDE. iii) Priming with rAd-E induced stronger CD4+ T cell responses which subsequently boosted with pDE to a greater extent than VV-E and rAd-E. Conclusion: These results indicate that priming with live viral vector vaccines could induce different patterns of E protein-specific CD4+ and CD8+ T cell responses which were significantly enhanced by booster vaccination with the DNA vaccine. Therefore, our observation will provide valuable information for the establishment of optimal prime-boost vaccination against DenV.

• 대한수의학회지
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• 제39권4호
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• pp.786-796
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• 1999

As an attempt to develop an effective control method against theileriosis, recombinant antigen protein was produced. Thirty-two kDa membrane protein(MP) gene of T sergenti was amplified through RT-PCR from extracted total RNA of T sergenti isolated in Chonbuk, Korea. The amplified 869 bp of Korean T sergenti membrane gene was cloned and the base sequences were analyzed. The amplified gene was cloned into E coli expression vector, pQE32 plasmid vector, and the vector was introduced into E coli strain M15 to produce the recombinant membrane protein. For the induction of T sergenti membrane protein(KTs-MP), the plasmid harboring E coli strain M15 were cultured in the presence of IPTG, and the recombinant protein were purified by $Ni^+$-NTA agarose. Then, to confirm the authenticity of the produced membrane protein, molecular weight of expressed recombinant KTs-MP was analyzed by SDS-PAGE and Western blotting. The molecular weight of expressed recombinant protein was 32 kDa as expected. The recombinant KTs-MP was successfully recognized by anti-His Tag antibody, antisera of T sergenti infected cattle and monoclonal antibody of T sergenti membrane protein. Therefore, we concluded that the authentic 32 kDa membrane protein of T sergenti was produced as immunologically recognizable form.

• BMB Reports
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• 제28권6호
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• pp.484-489
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• 1995

The product of bacteriophage T7 gene 2.5 is a single-stranded DNA binding protein and plays an important role in T7 DNA replication, recombination, and repair. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth (Kim and Richardson, 1993). The C-terminal truncated gene 2.5 protein ($GP2.5-{\Delta}21C$) cannot substitute for wild-type gene 2.5 protein in vivo; suggesting that the C-terminal domain of gene 2.5 protein is essential for protein-protein interactions (Kim and Richardson, 1994; J. Biol. Chem. 269, 5070-5078). Truncated gene 2.5 proteins lacking 19 residues ($GP2.5-{\Delta}19N$) and 39 residues ($GP2.5-{\Delta}39N$) from the amino-terminal domain were constructed by in vitro mutagenesis. $GP2.5-{\Delta}19N$ can support the growth of T7 phage lacking gene 2.5 while $GP2.5-{\Delta}39N$ cannot substitute for wild-type gene 2.5 protein in vivo; however, its ability to bind to single-stranded DNA is not affected. These results clearly demonstrate that the 20~39 amino-terminal region of gene 2.5 protein is required for T7 growth in vivo but may not be involved in DNA binding activity.

• 한국가금학회지
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• 제14권1호
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• pp.25-31
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• 1987

본 시험은 육계사료에 있어서 가장 경제적이고 효율적인 단백질 수준별 급여시기를 구명하여 육계사육의 생산성을 높이고자 실시하였다. 단백질 수준이 25, 23, 20 및 18%인 4가지 사료를 급여시기를 달리한 3개 처리구에 Aber Acres계 육계초생추 270수를 공시하여 1986년 6월 24일부터 1986년 8월 11일까지 7주에 걸쳐 시험을 실시한 결과를 요약하면 다음과 같다. 1. 증체량은 단백질 수준이 높아질수록 향상되었으며, T$_1$＞T$_2$＞T$_3$순으로 나타났다. (P〈0.05). 2. 사료중의 단백질 수준이 높을수록 사료섭취량이 증가하는 경향을 보였으며, 사료요구율은 T$_1$구와 T$_2$구는 비슷한 수준이었으며, T$_3$구가 가장 불량한 결과로 나타났으나 그 경향은 사료섭취량의 경우와 비슷하였다. 3. 단위 증체당 단백질 요구량은 고단백질 사료를 섭취할 때는 많았고 저단백질 사료를 섭취할때는 적었는데 일반적으로 체중이나 사료요구율과는 반비례하였고, 사료섭취량과는 정비례의 현상이 나타났다. 전기간에 걸친 단위 증체당 단백질 요구량은 T$_1$구와 T$_2$구 및 T$_3$구 간에 유의적인 차이를 보였다.(P〈0.05) 4. 육성율은 사료중의 단백질 수준에 영향을 받지 않았다. 5. 소득은 T$_1$구가 가장 높았으며 T$_2$구가 T$_3$구에 비하여 소득면에서 불량한 결과로 나타난 것은 고가의 고단백질 사료를 섭취하였으나 증체는 그에 따르지 못하는데 그 원인이 있는 것으로 보인다. 이상의 결과를 종합하여 볼 때 생산성 제고를 위하여 육계 사육 전반기에서의 고단백질 사료는 필수적인 것으로 보이며 가장 경제적이고 효율적인 사료중 단백질 수준별 급여시기는 T$_1$구가 적합할 것으로 사료된다.