• Title/Summary/Keyword: RFLP.

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Detection of Adulteration and Species Identification of Milk and Dairy Products using PCR: A Review (PCR을 이용한 품종동정 및 시유와 낙농제품의 진위판별 방법에 관한 연구: 총설)

  • Choi, Suk-Ho;Lee, Seung-Bae
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.4
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    • pp.253-262
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    • 2015
  • The authentication and implications of misleading labeling in milk and dairy products is important to protect against cheating consumers from adulteration and to alert sensitive consumers to any undeclared potential allergens. This need to support milk and dairy products labeling has led to the development of specific analytical techniques for the analysis of milk and dairy products ingredients. Recently, several methods based on polymerase chain reaction (PCR), including restriction fragment length polymorphism (PCR-RFLP), multiplex PCR, species-specific PCR, and real-time PCR, have been proposed as useful means for identifying species of origin in milk and dairy products, as well as quantifying and detecting any adulteration. These methods have particular advantages owing to their high specificity and sensitivity, as well as rapid processing time. In this review, we provide an updated and extensive overview of the PCR-based methods used for milk and dairy products authentication with a particular focus on the application of PCR methods to detect adulteration.

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Studies on the Specific Gene Expression in Lung Tissue of the Cloned Hanwoo (복제한우 폐조직에서 특이 유전자 발현에 관한 연구)

  • Kim, Sang-Hwan;Chung, Duck-Won;Lee, Ho-Jun;Hwang, Sue-Yun;Min, Kwan-Sik;Yoon, Jong-Taek
    • Reproductive and Developmental Biology
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    • v.33 no.1
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    • pp.19-24
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    • 2009
  • This study was conducted to investigate the specific expression genes in the cloned bovine tissues. Donor cells, cloned tissues were analysed by RAPD-RFLP method. The results were detected three genes (CH-U7B, CH-U7M and CH-U7P) in the cloned fetus. It was found a single copy genes by southern hybridization. Sequence analysis of CH-U7M gene was shown 99% homology to a previously reported EST from a cloned bovine fetus. The putative ORF was encode a protein of hydrophobicity index 0.03. Semi-quantitative RT-PCR by using the CH-LS001 specific primer was remarkably detected in the lung tissue of cloned fetus. Further investigation of these genes may provide one of the key information to explain the early death, abnormal fetus, large off-spring and the low pregnancy rate in the production of cloned bovine.

Association of Mahogany/Attractin Gene (ATRN) with Porcine Growth and Fat

  • Kim, Jong Bok;Zeng, Zhao Xun;Nam, Yu Jin;Kim, Younyoung;Yang, Shi Liu;Wu, Xiaolin;Lee, Chaeyoung
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.10
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    • pp.1383-1386
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    • 2005
  • Associations of restriction fragment length polymorphism in porcine attractin (ATRN) gene with average daily gain (ADG), live weight (LWT) and back fat thickness (BFT) were studied with one exotic composition population (Xianghuang) and three local breeds (Ningxing, Shaziling and Daweizi) in China. Two types of alleles were detected; one with the band size of 700 bp (A) and the other with the band size of 450 and 250 bp (B), and their genotypes were classified as AA, AB and BB. Polymorphisms were observed in the exotic breed, but not in the three local breeds. Individuals with the genotype BB showed larger ADG and LWT than those with AA in Xianghuang population (p<0.05). The breed specific effects of the porcine attractin gene in this study implied that identification of the polymorphisms and determination of genetic effects on phenotypes should be performed in the target populations.

The GSTP1 Ile105Val Polymorphism is not Associated with Susceptibility to Colorectal Cancer

  • Khabaz, Mohamad Nidal
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2949-2953
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    • 2012
  • The glutathione S transferase (GST) family is a major part of cellular defense mechanisms against endogenous and exogenous substances, many of which have carcinogenic potential. Alteration in the expression level or structure of the glutathione-S-transferase (GST) enzymes may lead to inadequate detoxification of potential carcinogens and consequently contribute to cancer development. A member of the glutathione-S-transferase (GST) family, GSTP1, is an attractive candidate for involvement in susceptibility to carcinogen-associated colorectal cancer. An $Ag{\rightarrow}G$ transition in exon 5 resulting in an Ile105Val amino acid substitution has been identified which alters catalytic efficiency. The present study investigated the possible impact of Ile105Val GSTP1 polymorphism on susceptibility to colorectal cancer. in Jordan We examined 90 tissue samples previously diagnosed with colorectal carcinoma, and 56 non-cancerous colon tissues. DNA was extracted from paraffin embedded tissues and the status of the GSTP1 polymorphism was determined using a polymerase chain reaction restriction fragment length polymorphism (RFLP) method. No statistically significant differences were found between colorectal cancer cases and controls for the GSTP1 Ile/Ile, Ile/Val and Val/Val genotypes. The glutathione S-transferase polymorphism was not associated with risk in colorectal cancer cases in Jordan stratified by age, sex, site, grade or tumor stage. In conclusion, the GSTP1 Ile105Val polymorphism is unlikely to affect the risk of colorectal cancer.

Polymorphism of p53 Gene Codon 72 in Endometrial Cancer: Correlation with Tumor Grade and Histological Type

  • Kafshdooz, Taiebeh;Tabrizi, Ali Dastranj;Ardabili, Seyyed Mojtaba Mohaddes;Kafshdooz, Leila;Ghojazadeh, Morteza;Gharesouran, Jalal;Abdii, Ali;Alizadeh, Hamideh
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.22
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    • pp.9603-9606
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    • 2014
  • Background: Endometrial cancer is the fourth most common cancer among women in developed countries. Patients with endometrial cancer may benefit from systemic chemotherapy alone or in combination with targeted therapies if the disease is clinically diagnosed prior to spread and metastasis to other organs. The aim of this study was to evaluate the prognostic role of p53 polymorphism and its correlation with tumor grade in human uterine endometrial carcinomas. Materials and Methods: A total of 75 patients with endometrial carcinomas were studied for possible mutations in exon 4 of the p53 gene using polymerase chain reaction and restricting fragment length polymorphism techniques and sequencing. Results: In recent study, The rate of homozygote genotype of pro/pro or Arg/Arg in high grade group was higher than in comparison with low grade one. In addition samples that were undigested in RFLP, showed mutation in exone 4. Conclusions: Our findings showed that high grade endometrial carcinomas are highly associated with TP53 polymorphisms in comparison with low grades.

MDR1 C3435T and C1236T Polymorphisms: Association with High-risk Childhood Acute Lymphoblastic Leukemia

  • Pongstaporn, Wanida;Pakakasama, Samart;Chaksangchaichote, Panee;Pongtheerat, Tanett;Hongeng, Suradej;Permitr, Songsak
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.7
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    • pp.2839-2843
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    • 2015
  • Background: MDR1, one of the most important drug-transporter genes, encodes P- glycoprotein (P-gp)-a transporter involved in protecting against xenobiotics and multi-drug resistance. The significance of the genetic background in childhood acute lymphoblastic leukemia (ALL) is not well understood. Materials and Methods: To evaluate whether C3435T and C1236T MDR1 polymorphisms are associated with the occurrence and outcome of ALL, 208 children with ALL (median age 5.0 yr) and 101 healthy Thai children were studied by polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) assay. Results: C3435T and C1236T MDR1 polymorphism are significantly associated with the high-risk group (OR= 2.6, 95%CI =1.164-5.808; P=0.028 and OR= 2.231, 95%CI =1.068-4.659; p=0.047, respectively), indicating that both may be candidates for molecular markers in the high-risk group of ALL.

Manganese Superoxide Dismutase (MnSOD Val-9Ala) Gene Polymorphism and Susceptibility to Gastric Cancer

  • Moradi, Mohammad-Taher;Yari, Kheirollah;Rahimi, Zohreh;Kazemi, Elham;Shahbazi, Mehrdad
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.2
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    • pp.485-488
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    • 2015
  • Background: Oxidative stress caused by the generation of reactive oxygen species plays an important role in human carcinogenesis. Manganese superoxide dismutase (MnSOD) Val-9Ala in the mitochondrial target sequence is the best known polymorphism of this enzyme. The purpose of the current research was to assess the association of MnSOD Val-9Ala genotypes with the risk of gastric cancer. Materials and Methods: This case-control study covered 54 gastric cancer patients compared to 100 cancer free subjects as controls. Extraction of DNA was performed on bioptic samples and genotypes were identified with a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Results: The frequencies of MnSOD Ala/Ala, Ala/Val and Val/Val genotypes in healthy individuals were 24.3, 66.7 and 9%, respectively. However, in gastric cancer patients, Ala/Ala, Ala/Val and Val/Val were observed in 24.0, 48.0 and 28.0% (p=0.01). In patients the frequency of MnSOD Val allele was higher (52%) compared to that in controls (42%). Conclusions: The results of this study show a positive association between MnSOD Val-9Ala gene polymorphism and risk of gastric cancer disease in Iranian population.

Identification and classification study of natural products by RAPD analysis (RAPD(Random Amplified Polymorphic DNA)법을 이용한 한약재의 판별 연구)

  • Kim, Dae-Weon;Kim, Do-Kyun;An, Sun-Kyong;Cho, Dong-Wuk
    • Korean Journal of Oriental Medicine
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    • v.3 no.1
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    • pp.153-167
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    • 1997
  • Conventionally, identification and classification methods of natural products include the morphological survey and assay of chemical disposition, sing these methods, however, is not satisfying for the precise identification of natural products because they are often valiable in the compositions and morphology To standardize the natural products identification and classification, genomic DNA analysis such as RAPD, RFLP and Amp-FLP can be adopted for this purpose. In this study, various ginsengs and bear gall bladder were tested for the development of genetic identification and classification method. Varieties of ginsengs such as, P. ginseng, P. quinquefolium, P. japonicus and P. notoginseng, were genetically analyzed by RAPD. Also, DNA isolated from Bear blood and gall bladder, Ursus thibetanus, Ursus americanus and Ursus arctos, were analyzed by the same method. The results demonstrated that the identification and classification of bear gall bladder and various ginsengs were possible by RAPD analysis. Therefore, this method was thought to be used as a additional method for the identification and classification of other natural products.

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Myeloperoxidase Polymorphism and Vitamin C Levels during Pregnancy Affect Maternal Oxidative Stress and Their Neonatal Birth Weights (임산부의 Myeloperoxidase 유전자다형성과 혈중 비타민 C 수준에 따른 모체의 산화 스트레스와 출생체중)

  • Park Bohyun;Kim Young-Ju;Park Eun Ae;Lee Hwayoung;Ha Eun-Hee;Park Jongsoon;Kim Jeongyoun;Hong Yun-Chul;Park Hyesook
    • Toxicological Research
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    • v.20 no.3
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    • pp.187-193
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    • 2004
  • This study aimed to determine the association of maternal oxidative stress and adverse pregnancy outcome with serum vitamin C concentration and a myeloperoxidase (MPO) genetic polymorphism during pregnancy. We investigated 450 pregnant women who visited Ewha Womans University Hospital for prenatal care during gestational weeks 24~28. During the second trimester, we measured serum vitamin C levels and urinary 8-hydroxyde-2'-deoxyguanosine (8-OHdG) and malondialdehyde (MDA) as an oxidative stress biomarker. We determined the presence of a maternal MPO polymorphism (G-to-A substitution at nucleotide 463) using a PCR-RFLP assay. We compared the level of oxidative stress and birth weight with the vitamin C concentration and the presence of the MPO polymorphism. The mean level of maternal oxidative stress tended to be higher and the birth weight lower for MPO type A/A than for types A/G and G/G. Vitamin C levels above the 75 percentiles were associated with reduced concentrations of urinary MDA and 8-OHdG but increased birth weight. Our data demonstrate that oxidative stress and neonatal birth weight are associated with the MPO genetic polymorphism, with the association modified by the maternal vita-min C levels.

Mutational Analysis of the MTHFR Gene in Breast Cancer Patients of Pakistani Population

  • Akram, Muhammad;Malik, Fa;Kayani, Mahmood Akhtar
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1599-1603
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    • 2012
  • Objectives: Since methylenetetrahydrofolate reductase (MTHFR) maintains the balance of circulating folate and methionine and blocks the formation of homocysteine, its regulation in relation to different cancers has extensively been studied in different populations. However, information on Pakistani breast cancer patients is lacking. The MTHFR gene has two most common mutations that are single nucleotide additions which result in change of amino acids C677T to Ala222val and A1298C to Glu429Ala. Methodology: 110 sporadic breast patients with no prior family history of cancer or any other type of genetic disorders along with 110 normal individuals were screened for mutations in exons 1 to exon 9 using single strand conformational polymorphism, RFLP and sequencing analyzer. Results: The p values for the 677CC, 677CT, and 677TT genotypes were 0.223, 0.006, and 0.077, respectively. Those for the 1298AA, 1298AC, and 1298CC genotypes were 0.555, 0.009, and 0.003, respectively. Conclusions: We found an overall a significant, weak inverse association between breast cancer risk and the 677TT genotype and an inverse association with the 1298C variant. These results for MTHFR polymorphism might be population specific in sporadic breast cancer affected patients but many other factors need to be excluded before making final conclusions including folate intake, population and disease heterogeneity.