• Journal of Applied Biological Chemistry
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• 제54권4호
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• pp.231-237
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• 2011

A gene encoding a putative alanine racemase in Xanthomonas. oryzae pv. oryzae was cloned, expressed and characterized. Expression of the cloned gene was performed in Escherichia coli BL21(DE3)pLys using a pET-21(a) vector harbouring $6{\times}histidine$ tag. Purification of the recombinant alanine racemase by affinity chromatography resulted in major one band by sodium dodecyl sulfate polyacryl amide gel electrophoresis analysis, showing about 45 kDa of molecular weight. The alanine racemase gene, cloned in this experiment, appears to be constitutively expressed in X. oryzae, as analyzed by reverse transcriptase polymerase chain reaction. The enzyme was the most active toward L-alanine and secondly D-alanine, showing a racemic reaction, thus the enzyme is considered as an alanine racemase. The enzyme was considerably activated by addition of pyridoxal-5-phosphate (PLP), showing that 75% increase in activity was observed at 0.3 mM, compared with control. D-Cysteine as well as L-cysteine significantly inhibited the enzyme activity. The inhibitions by cysteines were more prominent in the absence of PLP, showing 9 and 5% of control activity at 2 mM of addition, respectively. The enzyme was the most active at pH 8.0 and more stable at alkaline pHs than acidic pH condition.

• Asian-Australasian Journal of Animal Sciences
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• 제28권12호
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• pp.1774-1783
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• 2015

Milk lysozyme is the ubiquitous enzyme in milk of mammals. In this study, the cDNA sequence of a new chicken-type (c-type) milk lysozyme gene (YML), was cloned from yak mammary gland tissue. A 444 bp open reading frames, which encodes 148 amino acids (16.54 kDa) with a signal peptide of 18 amino acids, was sequenced. Further analysis indicated that the nucleic acid and amino acid sequences identities between yak and cow milk lysozyme were 89.04% and 80.41%, respectively. Recombinant yak milk lysozyme (rYML) was produced by Escherichia coli BL21 and Pichia pastoris X33. The highest lysozyme activity was detected for heterologous protein rYML5 (M = 1,864.24 U/mg, SD = 25.75) which was expressed in P. pastoris with expression vector $pPICZ{\alpha}A$ and it clearly inhibited growth of Staphylococcus aureus. Result of the YML gene expression using quantitative polymerase chain reaction showed that the YML gene was up-regulated to maximum at 30 day postpartum, that is, comparatively high YML can be found in initial milk production. The phylogenetic tree indicated that the amino acid sequence was similar to cow kidney lysozyme, which implied that the YML may have diverged from a different ancestor gene such as cow mammary glands. In our study, we suggest that YML be a new c-type lysozyme expressed in yak mammary glands that plays a role as host immunity.

• 농업과학연구
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• 제44권2호
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• pp.145-180
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• 2017

Alternanthera mosaic virus (AltMV) is a member of the genus Potexvirus which has been known for less than twenty years, and has been detected in Australasia, Europe, North and South America, and Asia. The natural host range to date includes species in at least twenty-four taxonomically diverse plant families, with species in at least four other families known to be infected experimentally. AltMV has been shown to differ from Potato virus X (PVX), the type member of the genus Potexvirus, in a number of ways, including the subcellular localization of the Triple Gene Block 3 (TGB3) protein and apparent absence of interactions between TGB3 and TGB2. Differences between AltMV variants have allowed identification of viral determinants of pathogenicity, and identification of residues involved in interactions with host proteins. Infectious clones of AltMV differing significantly in symptom severity and efficiency of RNA silencing suppression have been produced, suitable either for high level protein expression (with efficient RNA silencing suppression) or for Virus-Induced Gene Silencing (VIGS; with weaker RNA silencing suppression), demonstrating a range of utility not available with most other plant viral vectors. The difference in silencing suppression efficiency was shown to be due to a single amino acid residue substitution in TGB1, and to differences in subcellular localization of TGB1 to the nucleus and nucleolus. The current state of knowledge of AltMV biology, including host range, strain differentiation, host interactions, and utility as a plant viral vector for both protein expression and VIGS are summarized.

• 한국잠사곤충학회지
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• 제38권1호
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• pp.19-24
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• 1996

누에에서 생체 방어에 관련된 새로운 항 세균성 펩타이드 유전자를 탐색 분리하기 위하여 누에 체강에 비 병원성 세균인 Escherichia coli를 주사하여 면역 반응의 일환으로 발현량이 증가하는 유도 유전자 종류를 조사 하였다. 체강 주사 8시간 후 누에에서 cDNA 유전자 은행을 만들고, 정상 및 유도 누에에서 분리한 각각의 mRNA를 탐침으로 차별화 선별을 하였다. 차별화 선별 결과 정상보다도 유도 누에의 탐침을 사용한 막에서 강도가 높은 클론 32개를 선발하였고, 29개 클론에 대해 전체 또는 부분 염기 서열을 분석하여 DNA 상동성을 조사하였다. DNA 상동성 비교를 통해 생산한 발현 유전자 꼬리표 중에는 비교적 상동 유의성이 인정되어 그 실체를 추정할 수 있는 19개의 클론이 있었다. 특히 곤충의 면역 작용에 직접적으로 관계하는 항세균성 펩타이드 유전자, hemolin 유전자, transferrin 유전자 등 4종의 유전 자원을 확보할 수 있었다.

• 대한치과보철학회지
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• 제25권1호
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• pp.17-40
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• 1987

The purpose of this study was to clarify the effect of clasp design on abutment tooth adjacent to a distal extension base under the influence by the location of functional loading. The RPI clasp, the Akers clasp and the combination clasp were selected for evaluation. Tests were performed at 10Kg, 20Kg, 30Kg loads on the buccal, central, lingual, mesial and distal positions of loading platform of each mandibular distal extension partial denture. The laser reflexion method was used for three dimensional measurement of abutment movement, which is possible to measure precisely without contact. The movement in the mesiodistal(X), buccolingual(Y), and occlusoapical(Z) directions and the rotational movement(R) were measured, and in addition, the total movement (SV) as expressed by the three dimensional summation vector independent of direction was calculated. The data were analyzed using Student t-test, p<.05. The following results were obtained from this study; 1. Clasp design did not generally affect the direction of abutment tooth movement except the movement in an undesirable occlusal direction in case of the Akers clasp and the combination clasp. 2. The greater the load on the prosthesis, the greater was the abutment tooth movement, and the direction of abutment tooth movement was affected by positional loading. 3. Each prosthesis was dislodged from the test base under the small amount of load in the distal load position, and the buccal loading showed the greatest abutment tooth movement under the maximum load. 4. RPI clasp was evaluated as the most favorable design.

• Asian-Australasian Journal of Animal Sciences
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• 제29권9호
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• pp.1363-1370
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• 2016

Rennet, a complex of enzymes found in the stomachs of ruminants, is an important component for cheese production. In our study, we described that yak chymosin gene recombinant Pichia pastoris strain could serve as a novel source for rennet production. Yaks total RNA was extracted from the abomasum of an unweaned yak. The yak preprochymosin, prochymosin, and chymosin genes from total RNA were isolated using gene specific primers based on cattle chymosin gene sequence respectively and analyzed their expression pattern byreal time-polymerase chain reaction. The result showed that the chymosin gene expression level of the sucking yaks was 11.45 times higher than one of adult yaks and yak chymosin belongs to Bovidae family in phylogenetic analysis. To express each, the preprochymosin, prochymosin, and chymosin genes were ligated into the expression vector $pPICZ{\alpha}A$, respectively, and were expressed in Pichia pastoris X33. The results showed that all the recombinant clones of P. pastoris containing the preprochymosin, prochymosin or chymosin genes could produce the active form of recombinant chymosin into the culture supernatant. Heterologous expressed prochymosin (14.55 Soxhlet unit/mL) had the highest enzyme activity of the three expressed chymosin enzymes. Therefore, we suggest that the yak chymosin gene recombinant Pichia pastoris strain could provide an alternative source of rennet production.

• Asian-Australasian Journal of Animal Sciences
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• 제26권6호
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• pp.766-771
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• 2013

The purpose of this study was to investigate interaction effects of genes using a Harvester method. A sample of Korean cattle, Hanwoo (n = 476) was chosen from the National Livestock Research Institute of Korea that were sired by 50 Korean proven bulls. The steers were born between the spring of 1998 and the autumn of 2002 and reared under a progeny-testing program at the Daekwanryeong and Namwon branches of NLRI. The steers were slaughtered at approximately 24 months of age and carcass quality traits were measured. A SNP Harvester method was applied with a support vector machine (SVM) to detect significant SNPs in the CCDC158 gene and interaction effects between the SNPs that were associated with average daily gains, cold carcass weight, longissimus dorsi muscle area, and marbling scores. The statistical significance of the major SNP combinations was evaluated with $x^2$-statistics. The genotype combinations of three SNPs, g.34425+102 A>T(AA), g.4102636T>G(GT), and g.11614-19G>T(GG) had a greater effect than the rest of SNP combinations, e.g. 0.82 vs. 0.75 kg, 343 vs. 314 kg, 80.4 vs $74.7cm^2$, and 7.35 vs. 5.01, for the four respective traits (p<0.001). Also, the estimates were greater compared with single SNPs analyzed (the greatest estimates were 0.76 kg, 320 kg, $75.5cm^2$, and 5.31, respectively). This result suggests that the SNP Harvester method is a good option when multiple SNPs and interaction effects are tested. The significant SNPs could be applied to improve meat quality of Hanwoo via marker-assisted selection.

• 한국의학물리학회지:의학물리
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• 제17권4호
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• pp.207-211
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• 2006

kV cone-beam CT (CBCT)가 결합된 선형가속기의 등장으로 더욱 정확한 영상유도 방사선치료(Image-guided radiation therapy, IGRT)가 가능해졌다. IGRT는 영상장비를 이용해 표적의 이동을 보정할 수 있는 방사선치료기술로, CBCT를 이용한 IGRT의 경우 내부 장기의 정보를 바탕으로 병변의 이동을 정확히 알 수 있다. 내부 장기의 정보를 얻기 위해서는 방사선이 조사되기 바로 전에 환자의 CBCT 영상을 획득하여 치료계획 시 사용한 모의치료 CT영상과 정합을 수행하게 된다. 본 연구에서는 CBCT 영상의 재구성 품질에 따른 정합결과를 비교해 보았다. 6명의 환자로부터 총 56개의 CBCT 투과 정보를 획득하여 분석한 결과, 평행이동벡터의 차가 1 mm를 초과하는 경우는 단 3개에 불과하였다. 회전이동의 경우, x, y, z, 세 축을 모두 고려하였으며, 그 결과 총 168 ($56{\times}3$)개에서 단 3개만이 $1^{\circ}$ 이상의 차이를 나타냈다.

• Asian-Australasian Journal of Animal Sciences
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• 제17권1호
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• pp.146-152
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• 2004

Ghrelin is an acylated peptide recently identified as the endogenous ligand for the growth hormone (GH) secretagogues receptor 1a (GHS-R1a) and is involved in a novel system for regulating GH release. To understand the long-term effects of ghrelin, here we constructed six myogenic expression vectors containing the cDNA of swine mature ghrelin (pGEM-wt-sGhln, pGEM-wt-hGhln), ghrelin mutant of $Ser^3$ with $Trp^3$ (pGEM-mt-sGhln, pGEM-mt-hGhln) and truncated ghrelin derivative (pGEM-tmtsGhln, pGEM-tmt-hGhln) encompassing the first 7 residues of ghrelin (including $Ser^3$ substituted with $Trp^3$) and adding a basic amino acid, Lys (K) in the C-terminus. The constructs, pGEM-wt-sGhln, pGEM-mt-sGhln and pGEM-tmt-sGhln were linked with the ghrelin leader sequence, while the pGEM-wt-hGhln, pGEM-mt-hGhln and pGEM-tmt-hGhln were linked with a leader sequence from the human growth hormone releasing hormone (hGHRH). Intramuscular injection of 200 ${\mu}g$ pGEM-wt-sGhln or pGEM-tmt-sGhln augmented growth over 3 weeks in normal rats and peaked at day 21 or 14 post-injection respectively, whose body weight gains were on average approximately 6% or 19% heavier over controls. However, other injectable vectors had no such enhanced growth effects. Our results suggested that the efficacy of the ghrelin leader sequence was more effective than that of hGHRH in our system. Moreover, the results indicated that skeletal muscle might have the ability to posttranslationally modify the in vivo expressed ghrelin. And the most strikingly, the short ghrelin analog seems to mimic the biological effects more efficiently when compared with the full-length ghrelin.

• 대한치과교정학회지
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• 제50권2호
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• pp.75-85
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• 2020

Objective: The aim of this study was to evaluate changes in the nasal soft tissues, including movements of landmarks, changes in linear distances, and volumetric changes, using three-dimensional (3D) stereophotogrammetry after microimplant-assisted rapid palatal expansion (MARPE) in adult patients. Methods: Facial data were scanned using a white light scanner before and after MARPE in 30 patients. In total, 7 mm of expansion was achieved over a 4-week expansion period. We determined 10 soft tissue landmarks using reverse engineering software and measured 3D vector changes at those points. In addition, we calculated the distances between points to determine changes in the width of the nasal soft tissues. The volumetric change in the nose was also measured. Results: All landmarks except pronasale and subnasale showed statistically significant movement on the x-axis. Pronasale, subnasale, alar right, and alar left showed significant movement on the y-axis, while all landmarks except subnasale showed significant movement on the z-axis. The alar base width, alar width, and alar curvature width increased by 1.214, 0.932, and 0.987 mm, respectively. The average volumetric change was 993.33 ㎣, and the amount of increase relative to the average initial volume was 2.96%. Conclusions: The majority of soft tissue landmarks around the nasal region show significant positional changes after MARPE in adults. The nose tends to widen and move forward and downward. The post-treatment nasal volume may also exhibit a significant increase relative to the initial volume. Clinicians should thoroughly explain the anticipated changes to patients before MARPE initiation.