• Journal of Microbiology and Biotechnology
• /
• 제10권5호
• /
• pp.663-669
• /
• 2000

The fur (ferric uptake regulation) expression vector pMON2064 was modified to produce a Fur-fusion expression vector. A kinker site, factor Xa cleavage site, and several restriction endonuclease sites were introduced to facilitate easy cloning and isolating of the fusion protein. The resulting fusion expression vector, pMONSTER, was then used to make fusion expression vector, pMONSTER, was then used to make fusion proteins with $\beta$-galactosidase and the protease of the human immunodeficiency virus type 1 (HIV-1 PR). Strain SW4020 harboring the Fur $\beta$-galactosidase fusion vector produced blue colonies on a 5-bromo-4-chloro-3-indolyl-$\beta$-D-galactoside plate and the resulting 133 kDa fusion protein reacted with an anti-Fur antibody. The strain harboring the Fur-HIV-1 PR fusion vector produced a 29 kDa fusion protein, which also reacted with an anti-Fur antibody. The Fur-HIV-1 PR fusion protein was purified by a single column application that was designed to isolate the Fur protein. The purified Fur-HIV-1 PR fusion protein digested with factor Xa cleaved a recombinant Gag protein to release smaller fragments, including a p24 capsid protein. The Fur-HIV-1 PR fusion protein itself did not exhibit any proteolytic activity.

• Journal of Nutrition and Health
• /
• 제27권1호
• /
• pp.3-11
• /
• 1994

This study was carried out to examine how dietary protein and calcium levels in rats fed fat-enriched diet affect the total lipid and cholesterol contents of blood and tissues. Male Sprauge-Dawley rats weighing approximately 200g were fed six purified diets which contained 18%(w/w) beef tallow, 1% (w/w) cholesterol, two source of protein, casein or isolated soy protein (ISP) and three levels of dietary calcium, 0.1%, 0.4% and 1.0%, first, for four weeks, and second, for eight weeks. The contents of the total lipid, cholesterol and triglyceride in blood, liver, heart and feces were determined. After four weeks feeding serum lipid and cholesterol concentrations significantly decreased in rats fed 1.0% (w/w) level calcium, regardless of dietrary protein sources. After eight weeks, these concentrations were significantly lower in the rats fed soy protein than in casein-fed rats. As dietary calcium level increased serum and tissue lipid and cholesterol contents were decreased and fecal lipid excretion increased. It is concluded that hypolipidemic and/or hypocholesterolemic effects of soy protein and calcium were partly due to decrease in lipid absorption.

• 식품과학과 산업
• /
• 제52권1호
• /
• pp.60-67
• /
• 2019

Protein is a major nutrient of food and has long been studied for nutritional and utility value. Among them, rice protein is attracting attention because of its hypoallergenic characteristics and nutritional value. Rice proteins are divided into endosperm protein and bran protein depending on their location. The two proteins differ in their nutritional characteristics and applications. The endosperm protein is an insoluble protein and has an advantage of digestion and absorption. Rice bran protein dissolves well in water. Its amino acid value is high enough to be comparable to that of soy protein, and it has strong antioxidant ability. Rice protein is a healthy vegetable protein because of its health and hypoallergenic properties. It has been widely used in children's or patients' food, and recently for muscle supplement and health food. Rice protein is considered to be a very effective and useful material as it has been discovered so far.

• Journal of Microbiology and Biotechnology
• /
• 제24권4호
• /
• pp.568-576
• /
• 2014

TIM-1 (also known as KIM-1 and HAVcr-1) is a type I transmembrane glycoprotein member of the TIM family that may play important roles in innate and adaptive immune responses. The overexpression of proteins associated with membrane proteins is a major obstacle to overcome in studies of membrane protein structures and functions. In this study, we successfully coupled the overexpression of the TIM-1 protein with a C-terminal enhanced green fluorescent protein (GFP) tag in Escherichia coli. To the best of our knowledge, this report is the first to describe the overexpression of human TIM-1 in E. coli. The purified TIM-1-EGFP fusion protein recognized and bound directly to apoptotic cells and did not to bind to viable cells. Furthermore, we confirmed that the interactions of TIM-1-EGFP with apoptotic cells were blocked by TIM-1-Fc fusion proteins. This fusion protein represents a readily obtainable source of biologically active TIM-1 that may prove useful in future studies of human TIM-1.

• Animal cells and systems
• /
• 제1권2호
• /
• pp.317-321
• /
• 1997

Tau protein is one of the microtubule-associated proteins in the mammalian brain. In Alzheimer's disease, tau protein is immobilized in the somatodendritic compartment of certain nerve cells, where it forms a part of the paired helical filament (PHF). To understand the role of tau protein in the formation of PHF, a recombinant human tau protein expressed in Escherichia coli and five synthetic peptide fragments (peptide 1 to peptide 5), corresponding to the C-terminal region of tau protein, were prepared and their ability in self-assembly to form filamentous structures was examined. The recombinant human tau protein formed short rod-like structures in 0.1M MES buffer containing 1 mM $MgCI_2$, while a synthetic peptide fragment 1 containing 55 amino acid residues could assemble into a lot of long filamentous structures in water and particularly twisted helical structures in 0.1M MES buffer containing 1 mM $MgCI_2$. This suggests that the C-terminal region possesses a filament-forming ability and may be related to the formation of the helical structure by providing a powerful filament-forming driving force.

• Journal of Veterinary Science
• /
• 제22권4호
• /
• pp.49.1-49.6
• /
• 2021

The S1 protein of the infectious bronchitis virus (IBV) is a major structural protein that induces the production of the virus-neutralization antibodies. The monoclonal antibody against the IBV M41 S1 protein was used as a target for biopanning. After three rounds of biopanning, randomly selected phages bound to the monoclonal antibody. Sequence analysis showed that the dominant sequence was SFYDFEMQGFFI. Indirect competitive enzyme-linked immunosorbent assay showed that SFYDFEMQGFFI is a mimotope of the S1 protein that was predicted by PepSurf. The mimotope may provide information for further structural and functional analyses of the S1 protein.

• 한국동물위생학회지
• /
• 제27권4호
• /
• pp.355-369
• /
• 2004

The equine herpesvirus type 1 (EHV-1) immediate-early (IE) protein is a potent transactivator responsible for the activation of both early and late genes during the course of infection and is comprised of discrete functional domains that mediate its many functions. Interaction between trans activators such as the IE protein and various components of the RNA polymerase II transcription initiation machinery has been demonstrated to be critical for transactivation. In the present report, it is addressed the hypothesis that the IE protein interacts with various components of transcription machinery to mediate transactivation of target viral genes. In these studies, it is demonstrated that in vitro transcribed and translated IE protein interacts with TFIIB-agarose conjugate but not with TFIID-agarose conjugate. Additional immunoprecipitation studies using nuclear extracts derived from EHV-1 infected RK-13 cells confirmed that the IE protein interacts strongly with TFIIB, but fails to interact with TFIID. IR2, a truncated form of the IE protein lacking the potent transactivation domain and involved in the down-regulation of the IE gene, also interacted with TFIIB but not with TFIID. Studies were also performed to ascertain if particular TBP-associated factors (TAFs) could mediate IE or IR2 binding to TFIID. In vitro transcribed and translated TAF250 added to nuclear extracts generated from EHV-1 infected cells also failed to mediate an interaction between the IE protein or the IR2 protein and TFIID. This study demonstrated that the IE protein mediates transactivation of target viral genes by a mechanism that involves TFIIB. This is in contrast to mechanisms that have been proposed for both the herpes simplex virus ICP4 and VP16 protein which have been proposed to transactivate viral genes through interactions involving both TFIIB and TFIID. This study also intimates that IR2 mediate its repressive effects during the course of EHV-1 infection by a mechanism that involves sequestration of various transcription factors.

• Journal of Nutrition and Health
• /
• 제29권8호
• /
• pp.908-915
• /
• 1996

Protein concentration in human from 39 well-norished American women and its adequacy for growth of exclusively breastfed infants(BF) and breastfed infants fed supplementary foods(BFS)from 1-6 months postpartum were studied. Mean protein concentration of breast milk measured by Lowry et al., using human serum albumin as a standard, over the first 6 months lactation was 1.31$\pm$0.13g/dl. Concentration of protein was singnificantly higher at the first month of lactation (1.55$\pm$0.23g/dl)(P<0.05) than any other month studied. Mean volume of breast milk ranged from 662-848ml/day in the BE group and from 415-661ml/day in the BFS group during the first 6 months of lactation. Mean protein intake of infants ranged from 1.3-2.2g/kg in the BF group and from 1.4-2.1g/kg in the BFS group. Mean protein intake (g/kg body weight) of both BF and BFS groups was less than Recmmended Dietary Allowance(1989, USA) of 2.2g/kg except at 1 month of age. However, mean growth of the infants was normal according to NCHS reference, suggesting that the RDA for protein was unrealistically high for infants during 2-6 months of age. Protein provided by breast milk alone appeared adequate for normal growth during this time.

• Nutritional Sciences
• /
• 제1권1호
• /
• pp.3-7
• /
• 1998

The aim of the present investigation was to see whether an anabolic steroid, nandrolone phenylpropionate (NPP), exerts protienanabolic effects under such adverse nutritional conditions as protein deficiency and protein-energy malnutrition in male rats. feeding on a low-protein (8% casein) diet resulted in a marked reduction in body weight gain that was associated with reductions in body protein and protein content of gastrocnemius muscle. Administration of NPP (4 mg/kg body weight) did not alter muscle and body protein depletion induced by a low-protein diet. 50% food restriction caused reductions in body protein and in protein content of gastrocnemius muscle. These reductions were partially prevented by NPP (4 mg/kg body weight). Food restriction did not affect plasma concentration of corticosterone, insulin, or tetosterone plus dihydrotestosterone. On the other hand, neither plasma concentration of corticosterone nor insulin were affected by NPP. The present results show that anabolic steroids do not express anabolic effects under conditions of protein deficiency, but in protein-energy malnutrition, anabolic steroids exert their anabolic effects even in male rats.

• Toxicological Research
• /
• 제12권2호
• /
• pp.155-161
• /
• 1996

The enzymatic properties for NADPH-P450 reductase domain of a fusion protein between human cytochrome P450 1A1 and rat NADPH-P450 reductase expressed in Escherichia coli were investigated. The fusion plasmid pCW/1A1OR-expressed E. coli membrane showed high NADPH-cytochrome c reductase activity ($830.1\pm 85.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$), while pCW control vector and P 450 1A1 expression vector pCW/1A1 showed relatively quite low activity ($4.35\pm 0.49, 3.27\pm 0.50 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively). The kinetic curves for NADPH-cytochrome c reductase followed typical Michaelis-Menten kinetics. The $K_{max}$ and $V_{max}$ for NADPH-dependent reductase activity were $8.24\pm 2.61\mu $and $817.9\pm 60.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively, whereas those for cytochrome c-dependent reductase activity were $19.97\pm 2.86\mu M$ and $1303.5\pm 67.1 nmol\cdot min^{-1}\cdot mg protein^{-1}$. The reductase activities were also compared with those of rat, porcine and human liver microsomes. The activity of pCW/ 1A1OR-expressed E. coli membrane was 15.2-fold higher than that of rat liver microsome. Treatment with benzo(a)pyrene, 7-ethoxyresorufin and $\alpha$-naphthofiavone which are known as specific substrates or inhibitor for human P450 1A1 increased NADPH-cytochrome c reductase activity of fusion protein in E. coli membrane dose-dependently. These results demonstrate that the membrane topology of fused enzyme may be important for activity of its NADPH-P450 reductase domain.