• Asian-Australasian Journal of Animal Sciences
• /
• 제26권9호
• /
• pp.1218-1228
• /
• 2013

Fat quality is determined by the composition of fatty acids. Genetic relationships between this composition and single nucleotide polymorphisms (SNPs) in the stearoyl-CoA desaturase1 (SCD1) gene were examined using 513 Korean native cattle. Single and epistatic effects of 7 SNP genetic variations were investigated, and the multifactor dimensionality reduction (MDR) method was used to investigate gene interactions in terms of oleic acid (C18:1), mono-unsaturated fatty acids (MUFAs) and marbling score (MS). The g.6850+77 A>G and g.14047 C>T SNP interactions were identified as the statistically optimal combination (C18:1, MUFAs and MS permutation p-values were 0.000, 0.000 and 0.001 respectively) of two-way gene interactions. The interaction effects of g.6850+77 A>G, g.10213 T>C and g.14047 C>T reflected the highest training-balanced accuracy (63.76%, 64.70% and 61.85% respectively) and was better than the individual effects for C18:1, MUFAs and MS. In addition, the superior genotype groups were AATTCC, AGTTCC, GGTCCC, AGTCCT, GGCCCT and AGCCTT. These results suggest that the selected SNP combination of the SCD1 gene and superior genotype groups can provide useful inferences for the improvement of the fatty acid composition in Korean native cattle.

• BMB Reports
• /
• 제28권3호
• /
• pp.265-270
• /
• 1995

Acetolactate synthase (ALS, EC 4.1.3.18) is the first common enzyme in the biosynthesis of leucine, isoleucine, and valine. It is the target enzyme for several classes of herbicides, including the sulfonylureas, the imidazolinones, the triazolopyrimidines, the pyrimidyl-oxy-benzoates and the pyrimidyl-thio-benzens. The sulfonylurea-resistant ALS gene (SurB) from Nicotiana tabaccum [Lee et al. (1988) The EMBO J. 7, 1241-1248] was cloned into the bacterial expression plasmid pT7-7. The resulting recombinant plasmid pT7-ALS was used to transform an ALS-deficient Escherichia coli strain MF2000. MF2000 cells transformed with pT7-ALS grew in the absence of valine and isoleucine. ALS activities of 0.042 and 0.0002 ${\mu}mol/min/mg$ protein were observed in the crude extracts prepared from MF2000 cells transformed with plasmids pT7-ALS and pT7-7, respectively. In addition, the former crude extract containing mutant ALS was insensitive to inhibition by K11570, a new chemical class of herbicides. $IC_{50}$ values for K11570 were $0.13{\pm}0.01$ mM. For comparison, a plasmid pTATX containing the wild-type Arabidopsis thaliana ALS coding sequences was also expressed in MF2000. ALS activities of 0.037 ${\mu}mol/min/mg$ protein were observed, and the wild type ALS was sensitive to two different classes of herbicides, K11570 and ALLY, a sulfonylurea. $IC_{50}$ values for K11570 and ALLY were $0.63{\pm}0.07$ and $80{\pm}5.6$ nM, respectively. Thus, the results suggest that the sulfonylurea-resistant tobacco ALS was functionally expressed in the bacteria, and that K11570 herbicides bind to the regulatoty site of ALS enzymes.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권4호
• /
• pp.2433-2437
• /
• 2013

Vascular endothelial growth factor (VEGF) is a potent regulator of angiogenesis and thereby involved in the development and progression of solid tumours. Associations between three VEGF gene polymorphisms (-634 G/C, +936 C/T, and +1612 G/A) and breast cancer risk have been extensively studied, but the currently available results are inconclusive. Our aim was to investigate associations between three VEGF gene polymorphisms and breast cancer risk in Chinese Han patients. We performed a hospital-based case-control study including 680 female incident breast cancer patients and 680 female age-matched healthy control subjects. Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis was performed to detect the three VEGF gene polymorphisms. We observed that women carriers of +936 TT genotypes [odds ratio (OR) =0.46, 95% confidence interval (CI) = 0.28, 0.76; P=0.002] or 936 T-allele (OR=0.81, 95% CI= 0.68, 0.98; P=0.03) had a protective effect concerning the disease. Our study suggested that the +1612G/A polymorphism was unlikely to be associated with breast cancer risk. The -634CC genotype was significantly associated with high tumor aggressiveness [large tumor size (OR=2.63, 95% CI=1.15, 6.02; P=0.02) and high histologic grade (OR=1.47, 95% CI= 1.06, 2.03; P=0.02)]. The genotypes were not related with other tumor characteristics such as regional or distant metastasis, stage at diagnosis, or estrogen or progesterone receptor status. Our study revealed that the VEGF -634 G/C and +936 C/T gene polymorphisms may be associated with breast cancer in Chinese Han patients.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제35권1호
• /
• pp.7-13
• /
• 2017

In this study, changes in gene expression after administration of silk fibroin fragments ($size{\approx}30kDa$) were evaluated in MG63 cells using a cDNA microarray assay. In addition, the level of alkaline phosphatase (ALP) activity and cellular proliferation in the group administered moderately sized silk fibroin fragments ($size{\approx}30kDa$) (MSF) were compared to those in the group administered smaller silk fibroin fragments (size < 1 kDa) (SSF). The results of the cDNA microarray assay show increased expression of genes that are related to the cell cycle and inflammation. ALP, bone morphogenetic protein-7, bone morphogenetic protein receptor type IA, and runt-related transcription factor 2 exhibited significantly lower expression compared to control cells (fold ratio < 0.5). Relative ALP activity of the $100{\mu}g/mL$ MSF group was significantly lower than that of the SSF group (P < 0.05). Thus, the MSF group showed increased expression of genes associated with cellular proliferation and inflammation but decreased expression of genes associated with osteogenesis.

• Journal of Microbiology and Biotechnology
• /
• 제8권6호
• /
• pp.553-559
• /
• 1998

cDNA of human thrombopoietin (hTPO) amplified by polymerase chain reaction from a cDNA library of human fetal liver was cloned. EPO-like domains ($hTPO_{153} \;or\; hTPO_{l63})\; of\; hTPO(hTPO_{332}$) were expressed in Escherichin coli using several kinds of expression systems, such as ompA secretion, thioredoxin fusion, and the $P_L$ and T7 expression systems. To obtain $hTPO_{153}$ in soluble form, $hTPO_{153}$ cDNA was fused in-frame behind the gene encoding ompA signal sequence and thioredoxin protein. When fused with either of the genes, $hTPO_{153}$ was not expressed to the detectable level. However, a high level expression of the EPO-like domain of hTPO was obtained using the PL and T7 expression system. $hTPO_{153} \;or\; hTPO_{l63} cDNA were subcloned into the pLex and pET-28a(＋) vectors under the control of the inducible$ P_L\;T_7$ promoter, respectively. Proteins expressed using pl.ex vector and pET-28a(＋) detected in insoluble forms with an expression level of about 14% and 9% of total cellular proteins, respectively, and the level of expression was rapidly diminished in 2 h after the maximum level of expression was reached.

• Pediatric Gastroenterology, Hepatology & Nutrition
• /
• 제23권1호
• /
• pp.63-71
• /
• 2020

Purpose: Autoimmune hepatitis (AIH) is a chronic disease that may lead to cirrhosis. The immunopathogenesis of AIH is not fully understood and it mainly involves T-cell mediated mechanism. Macrophage migration inhibitory factor (MIF) is a pro-inflammatory cytokine that promotes T cell response and its polymorphism may serve as a severity marker of AIH. No previous study has considered investigating MIF polymorphism in children with AIH. Methods: Forty-two children with definite diagnosis of AIH were enrolled along with 100 age and sex matched controls. All participants were tested for polymorphism at -173GC (rs755622) of MIF gene. All patients received the standard protocol of steroid plus azathioprine to achieve remission. Liver biopsy was performed at time of diagnosis for all patients and only 18 of them underwent a second biopsy after treatment. Results: No statistically significant differences in the frequency of the genotypes GG and GC or in allele distribution were found in both patient and control groups (p=0.590, 0.640 respectively). Initial alanine aminotransferase (ALT) levels at the time of presentation was significantly higher in the GC group than GG group (p=0.020). GC genotype significantly correlated with disease relapse (r=0.41, p=0.007). Regression of necroinflammation and the fibrosis score in the second liver biopsy was statistically significant in the GG group (p<0.0001, p=0.010 respectively). Conclusion: MIF -173GC polymorphism is associated with clinically significant markers of pediatric AIH, including increased initial serum ALT levels, may help predict necroinflammatory/fibrosis regression effectively, following immunosuppressive treatment.

• 대한수의학회지
• /
• 제45권1호
• /
• pp.63-70
• /
• 2005

African swine fever (ASF) is an infectious disease of domestic and wild pigs for which there is no vaccine in the world. A proper surveillance of viral activity and a timely response to ASF outbreaks depend upon the rapid diagnosis of ASF viral infection. Internationally prescribed enzyme-linked immunosorbent assay (ELISA) is a fast, sensitive test routinely used in the diagnosis of the ASF. However, inactivated whole ASF virus antigen used in this test is a tedious to prepare and has a risk of outside exposure of infectious virus by laboratory accident during the preparation. An ASF virus noninfectious recombinant antigen is a safe and easily produced alternative antigen for use in diagnostic assay. We have cloned the ORF O61R gene of the ASF virus to generate a recombinant baculovirus producing the p12 protein in insect cells under control of the polyhedrin promoter as non-fusion protein. When used in an indirect ELISA, the p12 antigen showed reactivity with all known ASF positive pig sera but not with negative pig sera. Our results indicated that the p12 protein would be one of alternative antigens for diagnosis of the ASF.

• 미생물학회지
• /
• 제29권1호
• /
• pp.63-68
• /
• 1991

Several arsenic compound-resistant bacteria were isolated from Jung-Rang stream. The isolates, D-3, D-12, and D-14 were characterized phenotypically and genetically, and identified as Serratia liquefaciens, Klebsiella oxytoca, and Klebsiella pneumoniae, respectively. A plasmid of 67kb was found in Klebsiella oxytoca D-12 and designated as pMH12. Transfer of this plasmid from D-12 to E. coli HB101 was occurred, and the resulting transconjugant strains expressed the same level of heavy metal resistance as the donor strain. The physical presence of this plasmid in transconjugant was detected with agarose gel electrophoresis. Arsenite-sensitive derivatives of isolate D-12 were obtained with Mitomycin C treatment which cured pMH12. Antibiotics and heavy metal resistances were also examined to be used as a proper marker for the isolates in gene cloning. Isolate D-12 has resistance to several heavy metal ions such as $Cd^{2+}$ , $Zn^{2+}$ and $Hg^{ 2+}$ Also, all the other arsenite resistant isolates showed resistance to several heavy metal ions and antibiotics.

• Journal of Preventive Medicine and Public Health
• /
• 제31권1호
• /
• pp.15-26
• /
• 1998

p53 돌연변이 여부에 따른 유방암의 위험 인자를 찾아내고, p53 유전자 돌연변이가 유방암 발생에 관여하는 기전을 알아보기 위하여, 1993년 1월부터 1994년 11월 사이에 서울중앙병원에서 유방암으로 진단 받고 수술을 받은 환자 81명과, 이들과 연령, 거주 지역,교육 수준, 그리고 폐경 상태 등에 따라 1:1 혹은 1:2로 짝지은 대조군 121명을 대상으로, 임신력과 중요 영양소 및 총 칼로리 섭취량, 그리고 기타 유방암 관련 요인, p53 유전자 돌연변이 여부, 돌연변이 유형 등을 확인하여 다음과 같은 결과를 얻었다. 유방암 환자 81명중에 돌연변이가 관찰된 예는 25예(30.9%)였다. 이중에서 염기 치환이 20예(24.63%)로서 전이와 변위가 각각 10예(12.3%)였다. 전이 중에는 'C to T' 전이가 8예(9.9%), 'G to A'와 'A to G'는 각각 1예(1.2%)에 불과하였다. 변위는 'C to G'와 'G to T'가 각각 4예(4.9%)였고,'A to T'와 'T to A'가 각각 (1.2%)였다. 삽입과 탈락은 각각 4예(4.9%)와 1예(1.2%)였다. 전체 환자군과 전체 대조군 사이의 비교에서 분만횟수가 증가할수록 유방암의 대응비는 유의하게 감소하는 경향을 보였으며, 열량, 지방 및 단백질의 섭취량이 증가할수록 유방암의 위험도가 유의하게 증가하였다. p53 돌연변이 양성 환자군과 그 대조군 사이의 비교에서 지방과 단백질의 섭취량이 증가할수록 유방암의 위험도가 증가하였으나, 분만횟수증가는 유방암 위험도에 유의한 영향을 미치지 않았다. p53의 돌연변이가 확인되지 않은 경우에도 분만횟수 증가는 유의하지 않았으며, 식이요인 중에는 단백질만이 의미 있는 위험인자로 나타났다. 식이요인에 의한 유방암의 위험도 증가는 p53돌연변이가 존재하는 경우에 특히 두드러지게 나타났다. 이러한 결과는 식이 요인이나 그와 관련된 내분비적 요인에 의하여 p53 변이가 유발되고 이 변이가 유방 세포 암발생의 첫 단계로 작용하거나, 혹은 p53의 변이가 다른 유전자의 변이에 뒤 이어서 발생하는 촉진인자(promoter)로 작용할 수 있음을 시사하는 소견이다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권23호
• /
• pp.10225-10229
• /
• 2015

Genetic polymorphisms in homologous recombination repair genes cause an abnormal development of cancerous cells. In the present study we evaluated the possibility of breast cancer association with single nucleotide polymorphisms of RAD51, XRCC2 and XRCC3 genes. Polymorphisms selected in this study were RAD51 135G/C, XRCC2 Arg188His; and XRCC3 Thr241Met. Each polymorphism was genotyped using Polymerase chain reaction-restriction fragment length polymorphism in study cohort of 306 females (156 breast cancer patients and 150 controls). We observed that heterozygous variant genotype (GC) of RAD51 135 G/C polymorphism was associated with a significantly (OR=2.70; 95%CI (0.63-1.79); p<0.03) increased risk of breast cancer. In case of the XRCC3 gene we observed that frequency of heterozygous (OR=2.88; 95%CI (1.02-8.14); p<0.02) and homozygous (OR=1.46; 95%CI (0.89-2.40); p<0.04) genotype of Thr241Met polymorphism were significantly higher in breast cancer patients. For the Arg188His polymorphism of XRCC2, ~2fold increase in breast cancer risk (OR=1.6, 95%CI = 0.73-3.50) was associated with GA genotype with a p value for trend of 0.03. Our results suggest that the 135G/C polymorphism of the RAD51, Thr241Met polymorphism of XRCC3 and Arg188His polymorphism of XRCC2 can be independent markers of breast cancer risk in Pakistan.