• Title/Summary/Keyword: restriction fragment length polymorphism(RFLP)

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DNA Polymorphism of Insulin-like Growth Factor-binding Protein-3 Gene and Its Association with Cashmere Traits in Cashmere Goats

  • Liu, Haiying;Liu, Chao;Yang, Guiqin;Li, Hui;Dai, Jin;Cong, Yuyan;Li, Xuejian
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.11
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    • pp.1515-1520
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    • 2012
  • Insulin-like growth factor binding protein-3 (IGFBP-3) gene is important for regulation of growth and development in mammals. The present investigation was carried out to study DNA polymorphism by PCR-RFLP of IGFBP-3 gene and its effect on fibre traits of Chinese Inner Mongolian cashmere goats. The fibre traits data investigated were cashmere fibre diameter, combed cashmere weight, cashmere fibre length and guard hair length. Four hundred and forty-four animals were used to detect polymorphisms in the hircine IGFBP-3 gene. A 316-bp fragment of the IGFBP-3 gene in exon 2 was amplified and digested with HaeIII restriction enzyme. Three patterns of restriction fragments were observed in the populations. The frequency of AA, AB and BB genotypes was 0.58, 0.33 and 0.09 respectively. The allelic frequency of the A and B allele was 0.75 and 0.25 respectively. Nucleotide sequencing revealed a C>G transition in the exon 2 region of the IGFBP-3 gene resulting in R158G change which caused the polymorphism. Least squares analysis revealed a significant effect of genotypes on cashmere weight (p<0.0001), cashmere fibre length (p<0.001) and hair length (p<0.05) of the animals. The effect of genotypes on cashmere fibre diameter was not statistically significant (p>0.05). The animals of AB and BB genotypes showed higher cashmere weight, cashmere fibre length and hair length than the animals possessing AA genotype. These results suggested that polymorphisms in the hircine IGFBP-3 gene might be a potential molecular marker for cashmere weight in cashmere goats.

Analysis of Interleukin-6 Gene Polymorphisms in Behcet’s Disease Using RFLP and DHPLC (RFLP와 DHPLC를 이용한 베체트병 환자에서 Interleukin-6 유전자 다형성의 분석)

  • Jang, Won-Cheol;Park, Sang-Beom;Nam, Yun-Hyeong;Lee, Jae-Sik
    • Journal of the Korean Chemical Society
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    • v.50 no.2
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    • pp.123-128
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    • 2006
  • Behcet's disease (BD) is a systemic vasculitis characterized by recurrent oral and genital ulcers, and ocular inflammation, and which may involve the joints, skin, central nervous system and gastrointestinal tract. Although the exact pathogenesis for BD is not completely understood, it has been suggested that the disease is triggered in genetically susceptible individuals by environmental factors, such as microbial agents. It is noted that multiple genes, including MHC (major histocompatibility complex) and non-MHC genes, are implicated in the pathogenesis of BD. This study tries to determine whether IL-6 gene polymorphisms are associated with susceptibility to Behcet's disease in Koreans. Gene polymorphisms were typed by VNTR (variable number of tandem repeat), RFLP (restriction fragment length polymorphism), DHPLC (denaturing high performance liquid chromatography).There were no evidences for genetic association conferred by the IL-6prom polymorphism. However, significant differences in the IL-6vntr genotype and allele frequencies were found between patients with BD and controls. The IL-6vntr*C allele appeared to be an additional susceptibility gene to Korean BD. Further studies in other populations and gene are required to confirm these results.

Comparative Analysis of the Community of Culturable Bacteria Associated with Sponges, Spirastrella abata and Spirastrella panis by 16S rDNA-RFLP (16S rDNA-RFLP에 의한 Spirastrella abata와 Spirastrella panis 해면에 서식하는 배양가능한 공생세균 군집의 비교)

  • Cho, Hyun-Hee;Park, Jin-Sook
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.155-162
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    • 2009
  • A cultivation-based approach was employed to compare the culturable bacterial diversity associated with two phylogenetically closely related marine sponges, Spirastrella abata and Spirastrella panis, which have geologically overlapping distribution patterns. The bacteria associated with sponge were cultivated using MA medium supplemented with 3% sponge extracts. Community structures of the culturable bacteria of the two sponge species were analyzed with PCR-RFLP (restriction fragment length polymorphism) based on 16S rDNA sequences. The RFLP fingerprinting of 16S rDNA digested with HaeIII and MspI, revealed 24 independent RFLP types, in which 1-5 representative strains from each type were partially sequenced. The sequence analysis showed >98.4% similarity to known bacterial species in public databases. Overall, the microbial populations of two sponges investigated were found to be the members of the classes; Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria. The Alphaproteobacteria were predominant in the bacterial communities of the two sponges. Gammaproteobacteria represented 38.5% of bacterial community in S. abata. Whereas only 1.6% of this class was present in S. panis. Bacillus species were dominat in S. panis. Bacillus species were found to be 44.3% of bacterial species in S. panis, while they were only 9.7% in S. abata. It is interesting to note that Planococcus maritimus (8.1%, phylum Firmicutes) and Psychrobacter nivimaris (28.9%, phylum Gammaproteobacteria) were found only in S. abata. This result revealed that profiles of bacterial communities from the sponges with a close phylogenetic relationship were highly species-specific.

Mixed Infection of 16S rDNA I and V Groups of Phytoplasma in a Single Jujube Tree

  • Lee, Sang-Hun;Han, Sang-Sub;Cha, Byeong-Jin
    • The Plant Pathology Journal
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    • v.25 no.1
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    • pp.21-25
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    • 2009
  • Jujube trees infected with phytoplasma exhibit symptoms of typical witches' broom, such as yellowing, abnormally small leaves, short internodes and proliferation of shoots. A 1.2 kb fragment of the 16S rDNA from jujube phytoplasma was generated by R16F2n/R16R2 primer pair from earlier amplified P1/P7 PCR products of cloned jujube witches' broom phytoplasmas. Enzymatic restriction fragment length polymorphism (RFLP) and sequence analysis of 16S rDNA revealed that the jujube tree was infected with 16S rDNA I and V groups of phytoplasmas. Extensive comparative analyses of restriction enzyme profiles from Alu I, Hha I, Msp I, and Rsa I clearly classified the two into different phytoplasma groups. The phylogenie analyses based on 16S rDNA showed that the similarity of the two different clones was 87.5%. This is the first report of a mixed phytoplasmal infection in a single jujube tree.

Monitoring of Horizontal Gene Transfer from Agricultural Microorganisms to Soil Bacteria and Analysis of Microbial Community in Soils

  • Kim, Sung-Eun;Moon, Jae-Sun;Choi, Won-Sik;Lee, Sang-Han;Kim, Sung-Uk
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.563-566
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    • 2012
  • To investigate the possibility of horizontal gene transfer between agricultural microorganisms and soil microorganisms in the environment, Bacillus subtilis KB producing iturin and the PGPR recombinant strain Pseudomonas fluorescens MX1 were used as model microorganisms. The soil samples of cucumber or tomato plants cultivated in pots and the greenhouse for a six month period were investigated by PCR, real-time PCR, Southern hybridization, and terminal restriction fragment length polymorphism (T-RFLP) fingerprinting. Our data from Southern blotting and T-RFLP patterns suggest that the model bacteria do not give significant impacts on the other bacteria in the pots and greenhouse during cultivation.

Molecular Analysis of Exophiala Species Using Molecular Markers

  • Chee, Hee-Youn;Kim, Yoon-Kyoung
    • Mycobiology
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    • v.30 no.1
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    • pp.1-4
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    • 2002
  • Genetic relatedness of medically important Exophiala species such as E. dermatitidis, E. mansonii, and three E. jeanselmei varieties: jeanselmei, lecanii-corni, and heteromorpha was examined using PCR-RFLP(restriction fragment length polymorphism) of ribosomal DNA, M-13, $(GTG)_5$ and nucleotide sequences of ribosomal ITS(internal transcribed space) II regions. Three E. jeanselmei varieties showing distinct band patterns for each DNA markers as well as different nucleotide sequences of ribosomal ITS II regions could be considered as a separate species. E. dermatitidis and E. mansonii demonstrated the identical band patterns of RFLP of ribosomal DNA, M-13, and $(GTG)_5$ markers. However, nucleotides sequences of ribosomal ITS II region were different between these two species.

Lack of Association of a Common Polymorphism in the 3'-UTR of Interleukin 8 with Non Small Cell Lung Cancer in Kashmir

  • Bhat, Imtiyaz Ahmad;Pandith, Arshid A.;Bhat, Bashir A.;Naykoo, Niyaz A.;Qasim, Iqbal;Rasool, Roohi;Aziz, Sheik Aejaz;Shah, Zafar Amin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4403-4408
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    • 2013
  • Background: Chronic inflammation is considered as an important factor in the pathogenesis of lung cancer. The presence of inflammatory cells and higher levels of pro-inflammatory cytokines in the tumor microenvironment and their surrounding tissues is gaining much importance in research. Materials and Methods: One hundred ninety NSCLC cases and 200 age, smoking and sex matched controls were evaluated for association of IL-8 -251 (rs4073) and IL-8 -845 (rs2227532) in our population. Restriction fragment length polymorphism (RFLP) was used followed by direct sequencing for the detection of SNPs. Results: The IL-8 -845 polymorphism was not found in our population. No significant association was observed between the IL-8 -251 AT genotypes and IL-8 -25 AA genotypes and NSCLC (p=0.05) in our population. The IL-8 -251 A allele was also non-significant (p=0.05) in NSCLC patients. Conclusions: In conclusion, this report reveals lack of association between IL-8 - 251 A/T polymorphism and NSCLC in our Kashmir Valley population.

Detection of PSS Gene through Genomic DNA of Umbilical Cord Blood by PCR-RFLP in Piglets (자돈의 제대혈 Genomic DNA를 이용한 PSS 유전자 검색)

  • 김계웅;유재영;박홍양;윤종만;조규석;정재록;김건중;이종완
    • Korean Journal of Animal Reproduction
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    • v.27 no.2
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    • pp.97-102
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    • 2003
  • This study was carried out to find out PSS(Porcine Stress Syndrome) with the PSE(Pale, Soft, Exudative) in different piglets. These experiments were accomplished with the aid of PCR-RFLP(Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). The samples were collected and examined from umbilical cord blood of piglets of Yorkshire, Landrace and Crossbred. And then, the PCR products were digested by restriction enzyme, Hha I. The results obtained were as follows; The PCR products of the blood genomic DNA of ryanodine receptor gene were length of 1 .8kb in umbilical cord blood. Normal type(NN), heterozygous type(Nn) and recessively homozygous type(nn, PSS) as a result of digestion of restriction enzyme, Hha 1, were 90.0%, 10.0% and 0.0% in Yorkshire piglets, 76.2%, 19.0% and 4.8% in Landrace, 69.1%, 23.8% and 7.1% in crossbred, respectively. As already showing the above results, the blood from piglets umbilical cord can be availably used for the determination of genotypes of PSS because of easiness of blood collection without stress in live piglets.