scholarly journals Characterization of Ascaris from Ecuador and Zanzibar

2014 ◽  
Vol 89 (4) ◽  
pp. 512-515 ◽  
Author(s):  
A.M. Sparks ◽  
M. Betson ◽  
G. Oviedo ◽  
C. Sandoval ◽  
P.J. Cooper ◽  
...  
Keyword(s):  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Its Region ◽  
Restriction Enzymes ◽  
Zoonotic Transmission ◽  
Pcr Rflp ◽  
Double Digestion ◽  
Polymerase Chain ◽  
Shed Light

AbstractTo shed light on the epidemiology of ascariasis in Ecuador and Zanzibar, 177 adult worms retrieved by chemo-expulsion from either people or pigs were collected, measured and subjected to polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis of the ribosomal internal transcribed spacer (ITS) region. Upon double digestion with RsaI and HaeIII, PCR-RFLP analysis revealed the presence of A. lumbricoides in people and A. suum in pigs in Ecuador. In contrast, while there are no pigs on Zanzibar, of the 56 worms obtained from people, one was genotyped as A. suum. No additional genetic variation was detected upon further PCR-RFLP analysis with several other restriction enzymes. Upon measurement, worm mass and length differed by location and by species, A. suum being lighter and longer. While there is no evidence to suggest zoonotic transmission in Ecuador, an enduring historical signature of previous zoonotic transmission remains on Zanzibar.

Cloning, characterization and identification of polymorphism in TCR Zeta Gene in deoni cattle

2018 ◽  
Author(s):  
K. Swathi ◽  
M. Gnana Prakash ◽  
D. Sakaram ◽  
T. Raghunandan ◽  
A. Sarat Chandra ◽  
...  
Keyword(s):  
T Cell Receptor ◽  
Fragment Length Polymorphism ◽  
Bos Indicus ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Cell Receptor ◽  
Chain Reaction ◽  
Reading Frame ◽  
Pcr Rflp ◽  
Polymerase Chain

The cDNA encoding, T-cell receptor zeta (TCR z; CD247) molecule of Deoni cattle (Bos indicus), was isolated, cloned and sequenced in the present study. The CD247 cDNA comprised 1078 nucleotides including a 30 nucleotide 5¹-untranslated region (UTR), 495 nucleotide single open reading frame (ORF) and 553 nucleotide 3¹-UTR. Deduced amino acid of cattle CD247 sequence was two residues shorter than the corresponding sheep sequences. However, ruminant-specific insertions and substitutions in transmembrane (TM) and intra-cytoplasmic (IC) domain were present in cattle. Immunoreceptor tyrosine-based activation motifs (ITAMs), the important motifs for TCR signalling, were totally conserved among ruminants including cattle. The 3¹ - UTR region of the cattle CD247 was highly homologous to the corresponding region in the buffalo sequence and showed lack of polymorphism after polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using Hae III and Mse I restriction enzymes in cattle population. Phylogenetically, cattle sequence was closer to buffalo sequence under the ruminant’s lineage. The conserved nature of this gene ensures TCR integrity which is vital for induction of optimal and efficient immune response.

scholarly journals Identification of Pneumococcal Serotypes by PCR–Restriction Fragment Length Polymorphism

Diagnostics ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 196 ◽  
Author(s):  
García-Suárez ◽  
González-Rodríguez ◽  
Cima-Cabal ◽  
Yuste ◽  
Vazquez ◽  
...  
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Dna Sequences ◽  
Fragment Length ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Pcr Rflp ◽  
Restriction Fragment Length

Streptococcus pneumoniae shows more than 90 capsular serotypes that can be distinguished by their reactivity against antisera. The main objective of this work was the development of a molecular method for serotyping without the use of antisera. A computer program containing an algorithm was used to search in a database for potentially useful enzymes for Restriction Fragment Length Polymorphism-RFLP typing, in order to maximize the discrimination between different serotypes. DNA sequences of 90 serotypes for the region between dexB and aliA genes were compiled, and a computer screening of restriction enzymes was performed. The wzg–wzh–wzd–wze region and Sse9I restriction predicted unique PCR-RFLP patterns for 39 serotypes and eight serogroups. A second restriction enzyme resolved fragment specific patterns for 25 serotypes. The method was tested with 98 serotype-unknown clinical isolates. PCR-RFLP analysis deduced correct serotypes that were confirmed by Quellung reaction for 78.5% of the isolates.

scholarly journals Association of growth hormone (GH) gene polymorphism with growth and carcass in Sumba Ongole (SO) cattle

2017 ◽  
Vol 42 (3) ◽  
pp. 153 ◽  
Author(s):  
P. P. Agung ◽  
S. Anwar ◽  
W. P. B. Putra ◽  
M. S. A. Zein ◽  
A. S. Wulandari ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Gene Polymorphism ◽  
Fragment Length Polymorphism ◽  
Growth Parameters ◽  
Rflp Analysis ◽  
Cattle Breeding ◽  
Pcr Rflp ◽  
Gh Gene ◽  
Polymerase Chain ◽  
Dressing Percentage

A study was conducted to identify the polymorphism in the intron 3 of the Growth Hormone (GH) gene and also to evaluate the association of the GH gene polymorphism with growth parameters and dressing percentage in the Sumba Ongole (SO) cattle. A total of 267 individual DNA samples were used in the Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis. The SO cattle growth parameters data (n=44) including birth weight (BW), weaning weight at 205 days of age (WW205), yearling weight at 365 days of age (YW365) and also dressing percentage (DP) (n=122) were investigated in this study. There were three genotypes (AA, AB, and BB) of the GH gene based on the PCR-RFLP analysis with allele frequency was 0.87 and 0.13 for A allele and B allele respectively. The highest genotype frequency in the SO cattle is AA (0.76) and the lowest is BB (0.02). The Heterozygosity Observed (Ho) value in the SO cattle population is 0.23 and Polymorphism Information Content (PIC) value is 0.20. Therefore, the genetic diversity in the SO cattle based on the GH gene polymorphism is quite low. There is no association (P>0.05) in BW, WW205, YW365, and DP with genotypes of the GH gene. As the result, the GH gene in this study cannot be used as a genetic marker in the SO cattle breeding program.

A PCR-based approach to distinguish important Diadegma species (Hymenoptera: Ichneumonidae) associated with diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)

2004 ◽  
Vol 94 (5) ◽  
pp. 465-471 ◽  
Author(s):  
B. Wagener ◽  
A. Reineke ◽  
B. Löhr ◽  
C.P.W. Zebitz
Keyword(s):  
Plutella Xylostella ◽  
Fragment Length Polymorphism ◽  
Diamondback Moth ◽  
Restriction Enzymes ◽  
Morphological Characters ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Eastern And Southern Africa ◽  
Second Internal Transcribed Spacer ◽  
Cruciferous Plants

AbstractThe diamondback moth, Plutella xylostella (Linnaeus) has a cosmopolitan distribution and is one of the major pests on cruciferous plants. Biological control, especially with species of the genus Diadegma, has been successfully employed in several parts of the world, mainly in South East Asia. The taxonomy of this genus based on classical morphological characters is still unclear and misidentifications are reported. In the present study seven Diadegma species associated with P. xylostella were separated using polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) analyses. The second internal transcribed spacer (ITS2) of the ribosomal DNA (rDNA) was successfully amplified in all 167 individuals and digested using 11 different restriction enzymes. One restriction enzyme (CfoI) showed different restriction profiles in all species and also between two population samples of D. mollipla (Holmgren) from eastern and southern Africa. In addition, a new Diadegma species associated with P. xylostella from Ethiopia was discovered.

scholarly journals MMP-7 A-181G Polymorphism in Breast Cancer Patients from Western Iran

Breast Care ◽  
2015 ◽  
Vol 10 (6) ◽  
pp. 398-402 ◽  
Author(s):  
Kheirollah Yari ◽  
Ziba Rahimi ◽  
Mehrdad Payandeh ◽  
Zohreh Rahimi
Keyword(s):  
Breast Cancer ◽  
Cancer Patients ◽  
Fragment Length Polymorphism ◽  
Ductal Carcinoma ◽  
Lobular Carcinoma ◽  
Rflp Analysis ◽  
Breast Cancer Patients ◽  
Western Iran ◽  
Pcr Rflp ◽  
Polymerase Chain

Background: Matrix metalloproteinases (MMPs) are upregulated in tumors. The MMP-7 A-181G polymorphism is associated with increased expression of the MMP-7 gene. Aim of the present study was to investigate the association between the MMP-7 A-181G polymorphism and susceptibility to breast cancer. Patients and Methods: The MMP-7 A-181G variants were studied in a cohort of 251 subjects consisting of 100 breast cancer patients and 151 healthy controls; all were from Western Iran. The MMP-7 A-181G genotypes were identified using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Results: The frequencies of the MMP-7 AA, AG, and GG genotypes in healthy individuals were 34.4, 50.4, and 15.2%, respectively. In breast cancer patients, the frequencies of AA (34%), AG (52%), and GG (14%) genotypes (p = 0.95) were similar to those in the controls. There was a trend toward an increased frequency of the combined genotype of MMP-7 AG+GG in patients with lymph node metastasis (70.4%) compared to those without metastasis (66.7%). Also, in patients with invasive lobular carcinoma, the frequency of the MMP-7 AG+GG genotype tended to be higher (71.4%) compared to that in patients with invasive ductal carcinoma (66.2%) (p = 0.78). Conclusion: Our findings indicate that the MMP-7 A-181G polymorphism may not be correlated with susceptibility to breast cancer in our population.

scholarly journals Detection and characterization of Leptospira spp. isolated from aborted bovine clinical samples

2011 ◽  
Vol 81 (1) ◽  
pp. 21-25 ◽  
Author(s):  
Hassan Momtaz ◽  
Saadat Moshkelani
Keyword(s):  
Public Health Problem ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Clinical Samples ◽  
Dairy Herds ◽  
Important Public Health Problem ◽  
Pcr Rflp ◽  
Beef Herds ◽  
Bovine Abortion

Leptospira is recognized as an important public health problem worldwide, especially in tropical countries, and is a common cause of abortion in dairy and beef herds. The aim of the present study was to detect and characterize Leptospira as the causative agent of abortion in cattle using a PCR-RFLP in Chaharmahal va Bakhtiari and Isfahan provinces, Iran. A total of 220 bovine aborted foetuses and 120 vaginal discharges from an aborted calf were collected from 64 commercial dairy herds. After isolation of 60 Leptospira spp. from samples, RFLP analysis was carried out with HindIII and HaeIII restriction enzymes in reference strains and isolated for characterization. In a total of 340 specimens, 46 (20.9%) and 14 (11.66%) were identified positive for Leptospira spp. from aborted bovine foetuses and vaginal discharges, respectively. The present results also suggest that L. interrogans serovar hardjo has the highest prevalence in the region under study and L. hardjo is a major pathogen causing bovine abortion in Chaharmahal va Bakhtiari and Isfahan provinces of Iran.

scholarly journals Discrimination of Diploid Crucifer Species Using PCR-RFLP of Chloroplast DNA

HortScience ◽  
2004 ◽  
Vol 39 (7) ◽  
pp. 1575-1577 ◽  
Author(s):  
Claudia Cunha ◽  
Muhammet Tonguç ◽  
Phillip D. Griffiths
Keyword(s):  
Chloroplast Dna ◽  
Dna Sequences ◽  
Raphanus Sativus ◽  
Fragment Length Polymorphism ◽  
Diploid Species ◽  
Restriction Enzymes ◽  
Chain Reaction ◽  
Species Identity ◽  
Pcr Rflp ◽  
Polymerase Chain

Chloroplast DNA (cpDNA) was used to identify polymorphisms between crucifer species using the polymerase chain reaction-random fragment-length polymorphism (PCR-RFLP) technique. Ten primer pairs based on cpDNA gene sequences were used to amplify cpDNA fragments in Brassica oleracea L., B. rapa L., B. nigra (L.) Koch, B. napus L., B. carinata Braun, B. juncea (L.) Czern, and Raphanus sativus L. accessions. Amplified DNA sequences were then digested using 11 restriction enzymes to identify polymorphisms between the 7 species. Of the 110 combinations, 38 generated polymorphisms that discriminated one or more of the species. Genotyping of these polymorphisms in 10 accessions of each of the diploid species (B. oleracea, B. nigra, B. rapa and R. sativus) did not reveal segregating polymorphisms among accessions within species, indicating that they can be used to help determine species identity. Ten accessions of each of the amphidiploids B. napus, B. carinata and B. juncea were genotyped to infer their maternal ancestry. The diploid source of cpDNA in B. carinata was B. nigra in all accessions tested and B. rapa for nine of ten B. juncea accessions tested. Two B. napus accessions amplified polymorphisms shared with B. rapa, and eight accessions produced unique polymorphisms from neither B. rapa, B. oleracea or B. nigra. The polymorphisms identified in this study can be used to help confirm identity of the diploid crucifer species for taxonomic and conservation studies.

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