• Title/Summary/Keyword: Protein polymorphism

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Analysis of the spike glycoprotein gene and nonstructural protein gene of transmissible gastroenteritis virus using PCR and RFLP analysis (PCR과 RFLP분석을 이용한 transmissible gastroenteritis virus의 spike glycoprotein gene과 nonstructural protein gene의 분석)

  • Kwon, Hyuk-moo
    • Korean Journal of Veterinary Research
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    • v.36 no.3
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    • pp.627-633
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    • 1996
  • To analyze the genomic diversity of transmissible gastroenteritis virus (TGEV), the N-terminal half of the spike (S) glycoprotein gene and nonstructural protein gene (open reading frames 3 and 3-1) were amplified by reverse transcriptase reaction and polymerase chain reaction (RT-PCR), and analyzed using restriction fragment length polymorphism (RFLP) patterns of the amplified DNA. In this study, TGEV Miller (M6) and Purdue (P115) strains were used as reference strains, and two vaccine strains (MSV and STC3) and four Korea isolates (P44, VRI-WP, VRI-41, and VRI-48) were analyzed. All TGEV strains were amplified with three TGEV primer pairs. Although there was some exception in RFLP analysis, this method differentiated TGEV strains into following groups : Miller group (M6 and MSV), Purdue group (PUS, STC3, P44, VRI-WP, VRI-41, and VRI-48). Using Sau3AI and SspI, VRI-48 was differentiated from the Miller and Purdue type viruses. The RT/PCR in conjuction with RFLP analysis was a rapid and valuable tool for differentiating several strains of TGEV. This study revealed the occurences of distinct difference in genome of TGEV strains.

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The +1316 T/T Genotype in the Exon 3 of Uncoupling Protein Gene is Associated with Daily Percent Lay in Korean Native Chicken (한국 재래 닭의 Uncoupling Protein 유전자 Exon 3에서의 +1316 T/T 유전자형이 산란율에 미치는 효과 분석)

  • Oh J. D.;Lee J. H.;Hong Y. S.;Lee S. J.;Lee S. G.;Kong H. S.;Sang B. D.;Choi C. H.;Cho B. W.;Jeon G. J.;Lee H. K.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.239-244
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    • 2005
  • Uncoupling protein(UCP) is expressed exclusively in brown adipose tissue(BAT). It is blown to uncouple phosphorylation from oxidation and hence to be involved in energy metabolism and heat production, especially under cold exposure. In the present study, we identified single nucleotide polymorphism(SNP) in exon 3 of avUCP gene in Korean native chicken(KNC) population. It was detected a SNP T+1316C in exon 3 of avUCP gene by sequence analysis in KNC population. For PCR-RFLP analysis of the SNP T+1316C, used by AP III restriction enzyme. The result of PCR-RFLP analysis showed that allele T has two fragments of 255 bp and 86 bp, and allele C has only one fragment of 341 bp. The genotype frequencies were TT type, 0.7875; TC type, 0.1875 and CC type, 0.025; and the frequencies of allele T and C were 0.881 and 0.119, respectively in KNC population. Next study was conducted to investigate the effect of the SNP in avUCP gene on economic traits in the KNC population. The TT genotype had a significant higher daily percent lay(84.61) than CC genotype(p<0.05) in KNC population. This study may be useful for genetic studies of avCUP gene and selection on daily percent lay of KNC.

Polymorphic Characterization of Pharmaceutical Solids, Donepezil Hydrochloride, by 13C CP/MAS Solid-State Nuclear Magnetic Resonance Spectroscopy

  • Park, Tae-Joon;Ko, Dong-Hyun;Kim, Young-Ju;Kim, Yon-Gae
    • Bulletin of the Korean Chemical Society
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    • v.30 no.9
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    • pp.2007-2010
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    • 2009
  • Donepezil hydrochloride is a reversible acetylcholinesterase inhibitor that is used in the treatment of Alzheimer’s disease to improve the cognitive performance. It shows different crystalline forms including hydrates. Therefore, it is very important to confirm the polymorphic forms in the formulations of pharmaceutical materials because polymorphs of the same drug often exhibit significant differences in solubility, bioavailability, processability and physical/chemical stability. In this paper, four different forms of donepezil hydrochloride were prepared and characterized using X-ray powder diffraction, Fourier transform infrared, and solid-state nuclear magnetic resonance (NMR) spectroscopy. This study showed that solid-state NMR spectroscopy is a powerful technique for obtaining structural information and the polymorphology of pharmaceutical solids.

Identification of Luteovirus Nucleotide Sequences in Mild Yellow-Edge Diseased Strawberry Plants

  • Shaban Montasser, Magdy;Al-Awadhi, Husain;Hadidi, Ahmed
    • The Plant Pathology Journal
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    • v.18 no.1
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    • pp.1-5
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    • 2002
  • The availability of nucleotide sequences of the coat protein gene of Potato leafroll virus (PLRV) permitted the construction of DNA primers that were utilized for cDNA synthesis. Polymerase chain reaction (PCR) products of a 487 bp. and approximately 500 bp DNA fragments were amplified from nucleic acid extracts of PLRV-infected tissue and strawberry mild yellow-edge (SMYE) diseased strawberry tissue, respectively. The amplified DNA fragments were further differentiated by hybridization analysis with a CDNA probe for the coat protein gene of PLRV and restriction fragment length polymorphism (RFLP) analysis. These results suggest that a luteovirus is associated with the SMYE disease.

Bioinformatics Interpretation of Exome Sequencing: Blood Cancer

  • Kim, Jiwoong;Lee, Yun-Gyeong;Kim, Namshin
    • Genomics & Informatics
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    • v.11 no.1
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    • pp.24-33
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    • 2013
  • We had analyzed 10 exome sequencing data and single nucleotide polymorphism chips for blood cancer provided by the PGM21 (The National Project for Personalized Genomic Medicine) Award program. We had removed sample G06 because the pair is not correct and G10 because of possible contamination. In-house software somatic copy-number and heterozygosity alteration estimation (SCHALE) was used to detect one loss of heterozygosity region in G05. We had discovered 27 functionally important mutations. Network and pathway analyses gave us clues that NPM1, GATA2, and CEBPA were major driver genes. By comparing with previous somatic mutation profiles, we had concluded that the provided data originated from acute myeloid leukemia. Protein structure modeling showed that somatic mutations in IDH2, RASGEF1B, and MSH4 can affect protein structures.

Moleculay Cloning of the cDNA Encoding the 16 kDa Subunit of V-ATPase in Rat Brain (흰쥐 뇌에서 발현되는 16 kDa Vacuolar (H$^{+}$)-ATPase의 유전자 클로닝)

  • Shin, Song-Woo;Yoo, Min
    • Biomedical Science Letters
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    • v.6 no.3
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    • pp.165-170
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    • 2000
  • Vacuolar (H$^{+}$)-ATPase (V-ATPase) is an intracellular protein which consists of multiple subunits. It carries out acidification by pumping protons in the cell. This enzyme has also been found in the synaptic vesicles and may play an important role in the neurotransmission. We cloned cDNA fragments encoding the 16 kDa subunit of V-ATPase from the rat brain by RT-PCR and PCR using total RNA or recombinant phage DNA as templates. They contained the full coding sequences (468 bp) and one nucleotide at 3' region turned out to be different (A to C) when compared to the liver counterpart. However, this polymorphic difference did not cause any significant change in the primary structure of the protein because both GCA and GCC code for alanine. Our study would contribute to the understanding of the function of 16 M)a V-ATPase in the brain and of the mechanisms of neurotransmission.

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Platelet Derived Growth Factor-B and Human Epidermal Growth Factor Receptor-2 Polymorphisms in Gall Bladder Cancer

  • Mishra, Kumudesh;Behari, Anu;Kapoor, Vinay Kumar;Khan, M. Salman;Prakash, Swayam;Agrawal, Suraksha
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.14
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    • pp.5647-5654
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    • 2015
  • Gall bladder cancer (GBC) is a gastro-intestinal cancer with high prevalence among north Indian women. Platelet derived growth factor-B (PDGFB) and human epidermal growth factor receptor-2 (HER2) may play roles in the etiology of GBC through the inflammation-hyperplasia-dysplasia-carcinoma pathway. To study the association of PDGFB and HER2 polymorphisms with risk of GBC, 200 cases and 300 controls were considered. PDGFB +286A>G and +1135A>C polymorphisms were investigated with an amplification refractory mutation system and the HER2 $Ile^{655}Val$ polymorphism by restriction fragment length polymorphism. Significant risk associations for PDGFB +286 GG (OR=5.25) and PDGFB +1135 CC (OR=3.19) genotypes were observed for GBC. Gender wise stratification revealed susceptibility for recessive models of PDGFB +1135A>C (OR=3.00) and HER2 $Ile^{655}Val$ (OR=2.52) polymorphisms among female GBC cases. GBC cases with gall stones were predisposed to homozygous +286 GG and +1135 CC genotypes. Significant risk associations were found for ACIle (OR=1.48), GAVal (OR=1.70), GAIle (OR=2.00) haplotypes with GBC cases and GCIle haplotype with female GBC cases (OR=10.37, P=<0.0001). Pair-wise linkage disequilibrium revealed negative associations among variant alleles. On multi-dimensional reduction analysis, a three factor model revealed significant gene-gene interaction for PDGFB +286A>G, PDGFB +1135A>C and HER2 Ile165Val SNPs with GBC. Protein-protein interaction showed significant association of PDGFB and HER2 with the epidermal growth factor receptor signaling pathway.

Nucleotide Sequencing and PCR-RFLP of Insulin-like Growth Factor Binding Protein-3 Gene in Riverine Buffalo (Bubalus bubalis)

  • Padma, B.;Kumar, Pushpendra;Choudhary, V.;Dhara, S.K.;Mishra, A.;Bhattacharya, T.K.;Bhushan, B.;Sharma, Arjava
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.7
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    • pp.910-913
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    • 2004
  • Insulin-like growth factor binding protein-3 (IGFBP-3) gene is a structural gene associated with the growth and development of the animals. The present investigation was carried out to unravel nucleotide sequence and polymerase chain reactionrestriction fragment polymorphism (PCR-RFLP) of IGFBP-3 gene in buffalo. Genomic DNA was isolated from a total of 157 animals belonging to Murrah, Surti, Jaffarabadi and Nagpuri breeds of Indian riverine buffalo. A 655 bp of IGFBP-3 gene was amplified in all the breeds and amplicons were digested with Hae III, Taq I and Msp I restriction enzymes. On digestion with Hae III yielded single restriction pattern of 8 fragments of sizes 201, 165, 154, 56, 36, 19, 16 and 8 bp in all the animals studied. Similarly Taq I and Msp I also revealed single restriction pattern yielding fragments of sizes 240 and 415 bp and 145 and 510 bp, respectively. This shows nonpolymorphic nature of restriction sites in buffalo. Nucleotide sequencing of 587 bp of IGFBP-3 gene in Murrah buffalo was done and submitted to the GenBank (Accession No. AY304829). Nucleotide sequencing revealed an addition of 4 bases in the intronic region as compared to cattle.

Polymorphisms of Blood Proteins In Cheju Native Horses and Tsushima Native Horses (제주 재래마아 쓰시마 재래마의 혈액내 단백질의 다형)

  • 오유성;오문유;김세재;김기옥;고미희;모야박;양영훈
    • The Korean Journal of Zoology
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    • v.38 no.3
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    • pp.324-329
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    • 1995
  • The phylogenetic relationships between Cheju native horses and Tsushima native horses were studied by protein polymorphism analyses in 16 gene loci (Trypsin inhibitor: Ti, Chymotrypsin inhibitor: CTi, Albumin: Al, Esterase: Es, Transferrin: Tf, Hemoglobin: Hb, Catalase: Cat, Esterase D: EsD, Glutamate oxaloacetate transaminase: GOT, Glyoxalase I: GLO I, Acid phosphatase: AcP, Superoxide dismutase: SOD, Lactate dehydrogenase: LDH, Hexokinase: HK, Malate dehydrogenase: MDH, Malic enzyme: ME). All allelic patterns of the protein loci, except 5 loci (SOD, LDH, HK, MDH, ME), were polymorphic in both two populations. Gene frequencies of the polymorphic loci of the population of Cheju native horses were higher than those of Tsushima native horses. Average heterozygosity in Cheju native horses was 0.375, showing higher than that of Tsushima native horses (0.304). The Da distance and gene identity of two populations were 0.108 and 0.868, respectively. The phylogenetic tree constructed by these results and those previously reported in other horse populations, consisted of three clusters. From this phylogenetic tree, it could be suggested that Cheju native horses and Tsushima native horses had diverged from the Mongolian wild horse (Equus prsewolskii).

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Biochemical Characterization and Genetic Diversity of Pongamia pinnata (L.) Pierre in Eastern India

  • Kumari, Kanchan;Sinha, Amrita;Singh, Sanjay;Divakara, B.N.
    • Journal of Forest and Environmental Science
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    • v.29 no.3
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    • pp.200-210
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    • 2013
  • Biochemical characteristics of 24 Pongamia pinnata genotypes (candidate plus trees) from three agroclimatic zones were estimated and molecular characterization through RAPD markers was done. Various biochemical characters viz. seed oil, total carbohydrates, protein, acid value and Iodine number recorded significant variation among different genotypes. The highest seed oil content was 41.87% while seeds of 14 genotypes recorded above average (32.11%) for the trait. Seed oil and protein content exhibited a significant positive correlation and moderate heritability. Out of the initially selected twenty-five random primers, twenty-two RAPD primers were found to be highly reproducible and produced a total of 183 loci of which 147 (80.32%) loci were polymorphic. Percentage of polymorphism varied from 44% to 100% with an average of 80.62%. High level of genetic variation was found among different genotypes of P. pinnata. Both molecular and oil content (biochemical) markers appeared useful in analyzing the extent of genetic diversity in Pongamia and the result of these analyses will help to better understand the genetic diversity and relationship among populations. Overall, the Pongamia genotypes included in the study showed a correlation with their geographical origins such that genotypes from the same region tend to have higher genetic similarity as compared to those from different regions. However, in UPGMA based Nei's analysis, some genotypes were found not to be grouped based on geographical origins possibly due to the exchange of germplasm over time between farmers across the regions. The results from oil content analyses showed that several genotypes in 'Central and Western Plateau' agroclimatic zone of Jharkhand displayed a good potential for high oil content. The study provides insight about P. pinnata populations in Jharkhand (India) and constitutes a set of useful background information that can be used as a basis for future breeding strategy and improvement of the species.