• Title/Summary/Keyword: TaqI

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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.

Dopamine Transporter Gene and Dopamine D2, D3, D4 Receptor Gene Polymorphisms in Attention Deficit Hyperactivity Disorder (주의력결핍 과잉행동장애에서 도파민 전달체 및 도파민 D2, D3, D4 수용체 유전자 다형성)

  • Park, Pil-Sang;Kim, Dae-Kwang;Jung, Chul-Ho
    • Journal of the Korean Academy of Child and Adolescent Psychiatry
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    • v.19 no.1
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    • pp.19-27
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    • 2008
  • Objectives : The aim of this study was to examine the association of attention-deficit hyperactivity disorder (ADHD) in Korean populations with functional polymorphisms of six genes dopamine receptors (Ser311/Cys311 polymorphism, Taq1 A polymorphism, and Taq1 B polymorphism in DRD2, BalI polymorphism in DRD3, and promoter -521 C/T polymorphism and exon III 48 bp repeat polymorphism in DRD4) and one gene in dopamine transporter (DAT1). Methods : Participants were 58 children with ADHD and 110 control children. The genotypes were determined by PCR. Results : There was a statistically significant difference in genotype frequency of -521 C/T polymorphism within the promoter region of the DRD4 between two groups. Furthermore, in the male group, both genotype and allele frequencies showed statistically significant differences. Conclusion : Findings of the study indicate that -521 C/T polymorphism in promoter region of DRD4 appears to be a possible candidate gene for ADHD in Korean population.

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Genetic Diversity of Seven Strawberry mottle virus Isolates in Poland

  • Cieslinska, Miroslawa
    • The Plant Pathology Journal
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    • v.35 no.4
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    • pp.389-392
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    • 2019
  • The studies on detection of the Strawberry mottle virus (SMoV) have been conducted in Poland for breeding programme purpose and for producers of strawberry plant material. Leaf samples collected from infected strawberry plants were grafted on Fragaria sp. Indicators which were maintained in greenhouse for further study. Seven Fragaria vesca var. semperflorens 'Alpine' indicators infected by SMoV were used for the study aimed on molecular characterization of virus isolates. Partial RNA2 was amplified from total nucleic acids using the RT-PCR method. The obtained amplicons separately digested with BfaI, FauI, HaeIII, HincI, and TaqI enzymes showed different restriction profiles. The nucleotide sequences analysis of RNA2 fragment confirmed the genetic diversity of the SMoV isolates as their similarity ranged from 94.7 to 100%. Polish isolates shared 75.7-99.2% identity with sequence of the virus strains from the Czech Republic, the Netherlands, and Canada. Phylogenetic analysis resulted in grouping of the isolates found in Poland together with one of the Czech strain whereas two other from the Czech and the strains from the Netherlands and Canada created the separate cluster.

Associations between Vitamin D Receptor (VDR) Gene Polymorphisms and Colorectal Cancer Risk and Effect Modifications of Dietary Calcium and Vitamin D in a Japanese Population

  • Takeshige, Nobuyuki;Yin, Guang;Ohnaka, Keizo;Kono, Suminori;Ueki, Takashi;Tanaka, Masao;Maehara, Yoshihiko;Okamura, Takeshi;Ikejiri, Koji;Maekawa, Takafumi;Yasunami, Yohichi;Takenaka, Kenji;Ichimiya, Hitoshi;Terasaka, Reiji
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.5
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    • pp.2019-2026
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    • 2015
  • Much interest has been drawn to possible associations between vitamin D receptor (VDR) gene polymorphisms and colorectal cancer risk in conjunction with potentially protective effects of calcium and vitamin D. In a study of 685 cases of colorectal cancer and 778 community controls in Japan, we examined the associations of the FokI, BsmI, ApaI, and TaqI polymorphisms with colorectal cancer risk and effect modification by dietary calcium and vitamin D. Genotypes were determined by the PCR-RFLP method. The ApaI polymorphism seemed to be associated with a decreased risk of colorectal cancer, particularly of rectal cancer. The adjusted odds ratio of colorectal cancer for the ApaI AA and Aa genotypes combined versus the aa genotype was 0.83 (95% confidence interval [CI] 0.67-1.02), and the corresponding value for rectal cancer was 0.75 (95%CI 0.56-0.99). A decreased risk of colorectal cancer for the ApaI AA and Aa genotypes combined was more evident in individuals with high calcium intake (interaction p=0.055). The FokI polymorphism seemed to be associated with a decreased risk of colon cancer among those with high vitamin D intake (interaction p=0.09). The BsmI and TaqI polymorphisms were unrelated to colorectal cancer risk, and the null associations were not modified by calcium or vitamin D intake. In conclusion, the ApaI polymorphism may be associated with a decreased risk of colorectal cancer in Japanese, dependent on dietary calcium intake.

A riboprinting scheme for identification of unknown Acanthamoeba isolates at species level

  • Kong, Hyun-Hee;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.40 no.1
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    • pp.25-31
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    • 2002
  • We describe a riboprinting scheme for identification of unknown Acanthamoeba isolates at the species level. It involved the use of PCR-RFLP of small subunit ribosomal RNA gene (riboprint) of 24 reference strains by 4 kinds of restriction enzymes. Seven strains in morphological group I and III were identified at species level with their unique sizes of PCR product and riboprint type by Rsa 1. Unique RFCP of 17 strains in group II by Dde I. Taq I and Hae III were classified into: (1) four taxa that were identifiable at the species level. (2) a subgroup of 4 taxa and a pair of 2 taxi that were identical with each other. and (3) a species complex of 7 taxa assigned to A. castellanii complex that were closely related. These results were consistent with those obtained by 18s rDNA sequence analysis. This approach provides an alternative to the rDNA sequencing for rapid identification of a new clinical isolate or a large number of environmental isolates of Acanthamoeba.

Tobamoviruses of Green Peppers Growing on Hydroponic Systems (풋고추 수경재배에서 발생하는 tobamovirus의 특성)

  • Choi, Gug-Seoun;Kim, Jae-Hyun;Kim, Jeong-Soo;Kim, Hyun-Ran
    • Research in Plant Disease
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    • v.10 no.3
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    • pp.194-197
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    • 2004
  • Incidence of tobamovirus diseases was 100% at late growth stage of green pepper on hydroponic systems in plastic house. Infection frequency of the diseases showed 34% of Pepper mild mottle virus (PMMoV), 41.5% of Tobacco mild green mottle virus (TMGMV), and 24.5% of the co-infected viruses. The two viruses specifically reacted in DAS-ELISA prepared with each polyclonal antibody. A total of 77 pure tobamovirus isolates obtained from the crops was tested for pathotype determination. The isolation frequency of tobamovirus pathotype $P_{0}$ and $P_{1,2}$ was 61 % and 39%, respectively. All TMGMV isolates belonged to the pathotype $P_{0}$. In restriction enzyme analysis of the cDNAs synthesized with coat protein gene of PMMoV pathotype $P_{0}$ and $P_{1,2}$, the former had two TaqI sites but the later had one.one.

Identification of Deer Antler Species Using Sequence Analysis and PCR-RFLP of Mitochondrial DNA (사슴 미토콘드리아 DNA의 염기서열 및 PCR-RFLP분석에 의한 녹용의 종 감별)

  • Shin, Ki-Hyun;Shin, Sung-Chul;Chung, Ku-Young;Chung, Eui-Ryong
    • Food Science of Animal Resources
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    • v.28 no.3
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    • pp.276-282
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    • 2008
  • It is estimated that over 80% of deer antlers produced in the world are consumed in Korea. However, mislabeling or fraudulent replacement of costly antlers with cheaper ones is one of the most common problems in the domestic antler market. Therefore, there is a great need for the development of technology to identify species of antlers. This study was carried out to develop an accurate and reliable method for the identification and authentication of species or subspecies of antlers using DNA sequence analysis and comparison of mitochondrial cytochrome band D-loop region genes among antlers of five deer species, Cervus elaphus sibericus, Cervus elaphus canadensis, Cervus nippon, Cervus elaphus bactrianus and Rangifer tarandus. A variable region of cytochrome band D-loop genes was amplified using PCR with specifically designed primers and sequenced directly. The cytochrome band D-loop region genes showed different DNA sequences between the species of antlers and thus it is possible to differentiate between species on the basis of sequence variation. To distinguish between reindeer (Rangifer tarandus) antlers and other deer antlers, PCR amplicons of the cytochrome b gene were digested with the restriction enzymes NlaIV and TaqI, respectively, which generates a species-specific DNA profile of the reindeer. In addition, samples of 32 sliced antlers labeled Cervus elaphus sibericus from commercial markets were collected randomly and the mt DNA D-loop region of these antler samples was sequenced. Among the antler samples investigated, only 62.5% were from Cervus elaphus sibericus, and others were from Cervus elaphus bactrianus (25.0%), elk (Cervus elaphus canadensis) and reindeer (Rangifer tarandus). Our results suggest that DNA sequencing of mt DNA and PCR-RFLP methods using NlaIV and TaqI enzymes are useful for the identification and discrimination of deer antler species by routine analysis.

Genetic Relationships of Cattle Breeds Assessed by PCR-RFLP of the Bovine Mitochondrial DNA D-loop Region

  • Yoon, Du Hak;Lee, Hak Kyo;Oh, Sung Jung;Hong, Ki Chang;Jeon, Gwang Joo;Kong, Hong Sik;Lee, Jun Heon
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.10
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    • pp.1368-1374
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    • 2005
  • To investigate the genetic relationships among various cattle breeds, bovine mtDNA D-loop region was used in 411 animals of 18 cattle breeds, including 8 Asian Bos taurus, 7 European Bos taurus, 1 Asian Bos indicus, and 2 African Bos indicus. The size of amplified PCR products from mtDNA D-loop region was 964 bp and the products were digested by 15 different restriction enzymes. Two different band patterns were identified in eight restriction enzymes (BstXI, Hae III, Msp I, Apa I, Taq I, Alu I, BamH I, EcoN I) and the rest of restriction enzymes showed more than 3 different band patterns among which Apo I and MspR9 resulted in 7 different restriction patterns. The genotypes, number of haplotype, effective number of haplotype, and degree of heterozygosity were analyzed. Based on all the PCR-RFLP data, different haplotypes were constructed and analyzed for calculating genetic distances between these breeds using Nei's unbiased method and constructing a phylogenetic tree.

Relationship between Alcohol Withdrawal Symptoms and Dopaminergic Gene Polymorphisms(DRD2, DAT, COMT) in Alcohol Dependence Patients (알코올 의존 환자의 금단 증상에 영향을 미치는 도파민계(DRD2, DAT, COMT) 유전자 다형성)

  • Choi, Tai Young;Kim, Ho-Nam;Han, Doug-Hyun;Min, Kyung-Jun;Lee, Young-Sik;Na, Chul
    • Korean Journal of Biological Psychiatry
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    • v.13 no.3
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    • pp.178-190
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    • 2006
  • Objectives : We investigated the relationship of the alcohol withdrawal symptoms with genetic polymorphism among alcohol dependence patients. Method : The measuring instruments used in this study were the Clinical Institute Withdrawal Assessment for Alcohol(CIWA-Ar). We analyzed DRD2 TaqI A polymorphism, dopamine transporter(DAT 1) polymorphism, and catechol-O-methyltransferase(COMT) polymorphism in 108 male alcoholics and 76 healthy controls. Results : The major findings was as follows. No significant differences for genotype distribution or allele frequency were revealed comparing controls and alcoholic patients. DRD2 Taq I : The subscale score of auditory hallucination among CIWA-Ar scale in homozygote was significantly higher than in heterozygote(OR=1.34). The total score of CIWA-Ar scale in heterozygote was significantly higher than in homozygote. DAT1 : In the subject without DAT-9 gene allele, it was significantly higher of the subscale score of sweating, anxiety among CIWA-Ar scale than in the subject with DAT-9 gene allele. And The total score of CIWA-Ar scale in the subject without DAT-9 gene allele was significantly higher than in the subject with DAT-9 gene allele. COMT : The total score of CIWA-Ar scale in heterozygote was significantly higher than in homozygote. Conclusion : Our results suggest the relationship between specific genetic factors and the withdrawal symptoms of alcohol dependent patients. As the candidate gene of the severity of alcohol withdrawal syndrome, DRD2 Taq1 gene was recommended.

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Identification of Korean Native Goat Meat using Amplified Fragment Length Polymorphism (AFLP) DNA Markers (Amplified Fragment Length Polymorphism (AFLP) DNA Marker를 이용한 한국 재래흑염소육 감별)

  • 정의룡
    • Food Science of Animal Resources
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    • v.22 no.4
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    • pp.301-309
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    • 2002
  • This study was carried out to develop the breed-specific DNA markers for breed identification of Korean native goat meat using amplified fragment length polymorphism (AFLP)-PCR techniques. The genomic DNAs of Korean native goat, imported black goat and four dairy goat breeds(Saanen, Alpine, Nubian and Toggenburg) were extracted from muscle tissues or blood. Genomic DNA was digested with a particular combination of two restriction enzymes with 4 base(Mse I and Taq I) and 6 base(EcoR I and Hind III) recognition sites, ligated to restriction specific adapters and amplified using the selective primer combinations. In AFLP profiles of polyacrylamide gels, the number of scorable bands produced per primer combination varied from 36 to 74, with an average of 55.5. A total of 555 bands were produced, 149(26.8%) bands of which were polymorphic. Among the ten primer combinations, two bands with 2.01 and 1.26 kb in M13/H13 primer and one band with 1.65 kb in E35/H14 primer were found to be breed-specific AFLP markers in Korean native goat when DNA bands were compared among the goat breeds. In the E35/H14 primer combination, 2.19, 2.03, 0.96 and 0.87 kb bands detected in imported black goat, 2.13 kb band in Saanen breed and 2.08 kb band in Nubian breed were observed as breed-specific bands showing differences between goat breeds, respectively. The E35/H14 primer combination produced four DNA bands distinguished between Korean native goat and Saanen breed. The is study suggested that the breed specific AFLP bands could be used as DNA markers for the identification of Korean native goat meat from imported black goat and dairy goat meats.