• BMB Reports
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• 제28권4호
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• pp.353-358
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• 1995

Epitope tagging is the process of fusing a set of amino acid residues that are recognized as an antigenic determinant to a protein of interest. Tagging a protein with an epitope facilitates various immunochemical analyses of the tagged protein with a specific monoclonal antibody. The monoclonal antibody H8 has subtype specificity for an epitope derived from the preS2 region of hepatitis B virus surface antigen. Previous studies on serial deletions of the preS2 region indicated that the preS2 epitope was located in amino acid residues 130~142. To test whether the amino acid sequence in this interval is sufficient to confer on proteins the antigenicity recognizable by the antibody H8, the set of amino acid residues in the interval was tagged to the amino terminal of ${\beta}$-galactosidase and to the carboxyl terminal of the truncated $p56^{lck}$ fragment. The tagged ${\beta}$-galactosidase, expressed in Escherichia coli, maintained the enzymatic activity and was immunoprecipitated efficiently with H8. The tagged $p56^{lck}$ fragment, synthesized in an in vitro translation system, was also immunoprecipitated specifically with H8. These results demonstrate that the amino acid sequence of the preS2 region can be used efficiently for the epitope tagging approach.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제30권3호
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• pp.186-192
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• 2004

Purpose: Human tooth proteins are highly heterogeneous, comprising diverse proteins derived from a number of genes. The attempts to identify protein for activity of tooth matrix proteins have been defied by several factors. First, the amount of proteins within teeth is very small relative to many extracellular matrix proteins of other tissues. Second, the bioassay system is tedious and needed for long time. Therefore we tried to find easy techniques, which increase the product rate, and an assay of small proteins, with which amino acid sequence is possible without additional procedures. Materials and Methods: Total protein were extracted from 300 g enamel removed teeth and 600 g teeth with 4 mol/L guanidine HCl and purified by gel chromatography. Aliquot of proteins was implanted into muscle pouches in Sprague-Dawley rats for bioassay. By SDS-PAGE and membrane blotting, molecular weight of each protein was estimated and a partial amino acid sequence was obtained. Each fraction blotted on the membrane was cut out and inserted in rat ectopic model. Results: In dissociative method, total tooth proteins were obtained 1mg/ml from enamel removed teeth and 3.5 mg/ml from teeth. In SDS-PAGE, four clear bands at the sites corresponding to 66, 40, 20 and 18 kD. Especially The 66 kD band was clearly exhibited. Amino acid sequencing from tooth could be possible using PVDF membrane blotting technique. In amino acid sequencing, 66 kD protein was identified as albumin. Conclusion: Compared with conventional method for extraction of teeth protein and bioassay of proteins, the methods in this study were easy, time-saving and more productive technique. The matured tooth proteins omitting additional procedure of mechanical removal of enamel were simply analyzed using blotted PVDF membrane. This method seems to make a contribution as a technique for bioassay and amino acid sequencing of protein.

• Journal of Nutrition and Health
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• 제7권4호
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• pp.12-20
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• 1974

Quantitative analysis was achieved by gas-liquid chromatographic method (GLC) with a single column system of OV-17 for 16 of protein amino acids in mushroom (agaricus bisporus). The quantities of protein amino acids in mushroom were determined $48.32{\sim}255.94mg%$ alanine, $108.6F{\sim}364.82mg%$ glycine, $124.30{\sim}314.17mg%$ Valine, $32.99{\sim}418.79mg%$ leucine and isoleucine, $151.78{\sim}669.07mg%$ threonine, $88.12{\sim}4F6.3Fmg%$ Serine, $21.9F{\sim}114.94mg%$ Hydroxyproline, $20.F4{\sim}174.63mg%$ proline, $34.52{\sim}173.59mg%$ Methionine, $225.25{\sim}1417.61mg%$ Aspartic Acid, $10F.00{\sim}392.17mg%$ Phenylalanine, $12F.46{\sim}535.65mg%$ Glutamic Acid and Lysine, Tyrosine in trace amount.

• Journal of Nutrition and Health
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• 제29권7호
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• pp.689-702
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• 1996

The purpose of this experiment was to compare the effects of tryptophan administration on nutritional status of female rats which consumed reserpine and 6% casein diet with different carbohydrate contents(87%, 65%, 44% respective). Final body weight, body weight gain, FER, plasma amino acid concentration and microsomal cytochrome P 450 content in liver were measured and microscopic structure of hepatocytes was observed. In low-protein diet, the higher the carbohydrate content of diet was, the lower the damage was in the rat's liver. Tryptophan administration after dose of reserpine induced more effective recovery from liver damage of rats in high carbohydrate diet group than that in low carbohydrate diet group. In conclusion, the general nutritional assessments such as final body weight and body weight gain provided better estimate of the degree of structural changes in hepatocytes than functional assessment such as plasma amino acid concentration or liver microsomal cytochrome P450.

• 약학회지
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• 제39권6호
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• pp.676-680
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• 1995

The nucleotide sequence of Xbal-Mbol fragment of pKH7, a chloramphenicol-resistant($Cm^{r}$) plasmid isolated from multidrug-resistant S. aureus SA2, has been determined. Xbal-Mbol fragment of pKH7 was found to contain two ORFs. One ORF encoded Rap and the other encoded CAT protein. The deduced amino acid sequences of Rep and CAT of pKH7 were compared to those of pUB112 and pC221. Comparisons revealed that there was one amino acid difference in CAT between pKH7 and pUB112. CAT of pKH7 exhibited 98.6% amino acid identity to that of pC221. In case of Rep proteins, a slightly lower homology of 96.4% and 86.7% in amino acid sequences was observed between pKH7 and pUB112 and between pKH7 and pC221, respectively.

• 한국식품영양학회지
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• 제14권6호
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• pp.562-567
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• 2001

In order to utilize okara protein as a food auditive, nutritional composition of soymilk okara was investigated. Protein in okara Is highly insoluble due to excessive heat treatment during soymilk processing. Protein content of okara was 37.3% as compared to 42.5 % for soybean. Carbohydrate and lipid contents of okara were 40.6% and 17.9%, respectively. Okara lipid extracted with chloroform-methanol consisted of neutral lipid, glycolipid and phospholipid, with neutral lipid making up 98.6% . Linoleic acid, ileic acid, and palmitic acids accounted for about 80% of the total fatty acids with linoleic acid sharing 50.3% of the total. Amino acid composition of okara protein was dissimilar to that of soy Protein : Cysteine was totally absent in okara while lysine, which is the limiting amino acid of soy protein, was present in higher amount in okara on dry weight basis. Both aqueous extract of okara protein and soy Protein were found to have ACE inhibitory activity.

• 한국식품영양학회지
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• 제30권4호
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• pp.796-803
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• 2017

This study investigated the crude protein and amino acid contents of local agricultural products widely and specifically grown in Korea, including 25 vegetables and 13 fruits. The crude protein content of vegetables and fruits ranged from 0.46 to 6.53% and 0.29 to 2.23%, respectively. Totally, 17 types of amino acids were found in most samples. The total amino acid content of vegetables and fruits ranged from 457.38 to 9,303.18 mg% and 368.82 to 3,118.75 mg%, respectively. The total amino acid contents of garlic and passion fruit was higher compared to other vagetables and fruits. The calibration curves of the standard components showed good linearity ($r^2$>0.99), except Met ($r^2=0.989$). The limits of LOD and LOQ were in the range 0.034 to $0.991{\mu}g/mL$ and 0.009 to $0.474{\mu}g/mL$, respectively. The results of the study can serve as a fundamental source of information regarding crude protein and amino acids contents in food, for diet planning.

• Journal of Microbiology
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• 제33권2호
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• pp.165-170
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• 1995

The 356 amino acid long lambda integrase protein of bacteriophage .lambda. constains two autonomous DNA binding domains with distinct sequence specificities. The amino terminal domain of integrase is implicated to bind to the arm-type sequences and the carboxyl domain interacts with the coretype sequencess. As a first step to understand the molecular mechanism of the integrase-DNA interaction at the arm-type site, the int(am)94 gene carrying an amber mutation at the 94th codon of the int was cloned under the control of the P$\_$tac/ promoter and the lacI$\_$q/ gene. The Int(am)94 mutant protein of amino terminal 93 amino acid residues can be produced at high level from a suppressor free strain harboring the plasmid pInt(am)94. The arm-type binding activity of Int(am)94 were measured in vivo and in vitro. A comparison of the arm-type binding properties of the wild-type integrase and the truncated Int(am)94 mutant indicated that the truncated fragment containing 93 amino acid residues carry all the determinants for DNA binding at the arm-type sites.

• Journal of Plant Biology
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• 제16권1_2호
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• pp.30-38
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• 1973

To study the response to plant growth by the environmental factors, the effects of application of nitrogen on changes in the yield, crude protein, amino acids, chlorophyll, carotene, total phosphorus, acid-soluble phosphorus, phospholipids, RNA, and DNA were investigated with westerworlds 9Lolium sublatum) and perennial rye-grasses (Lolium perenne). The amounts of dry weight, crude protein, amino acids, chlorophyll, carotene, total phosphorus, acid-soluble phosphorus, phospholipids, RNA and DNA of both rye-grasses increased with adequately increasing nitrogen, and reached a maximum with an adequate application of nitrogen. The relationships between yields and crude protein contents, crude protein and RNA contents, and yields and RNA contents of westerworlds and perennial rye-grasses were found to be positively correlated, respectively. Therefore, in general, the response to plant growth by the environmental factors such as nitrogen nutrient may be summarized as follows: Environmental factors\longrightarrowDNA\longrightarrowRNA\longrightarrowProtein\longrightarrowPlant growth

• 한국식품과학회지
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• 제30권6호
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• pp.1388-1393
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• 1998

젖산균 및 효모를 이용하여 전통안동식혜를 제조하여 저장하는 동안 질소화합물 및 아미노산의 변화를 조사하였다. 숙성기간동안 조단백질의 함량은 주발효 기간인 4일까지는 증가하였으며, 아미노태질소는 시간이 경과할수록 증가하였으며 2일째 37.50 mg였으며 이때 식혜의 맛이 가장 좋았다. 수용성 및 염용성 단백질은 시간이 경과함에 따라 점차 감소하였고 주요 아미노산은 proline 및 aspartic acid였으며 methionine은 시간이 경과함에 따라 점차 증가하였으나 lysine은 감소하였다. 수용성 및 염용성 단백질의 아미노산 조성은 총 17종이었고, 이 중 glutamine acid 및 aspartic acid의 함량이 가장 많았다. 염용해성 단백질의 경우 arginine은 시간이 경과함에 따라 점차 증가하였다.