scholarly journals Analysis of Kinetoplast DNA from Mexican Isolates ofLeishmania (L.) mexicana

2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Omar Hernández-Montes ◽  
Saúl González Guzmán ◽  
Federico Martínez Gómez ◽  
Douglas C. Barker ◽  
Amalia Monroy-Ostria
Keyword(s):  
Rflp Analysis ◽  
Restriction Enzymes ◽  
The Other ◽  
Polymorphism Analysis ◽  
Specific Primers ◽  
Restriction Fragment Polymorphism ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Similar Restriction ◽  
Polymerase Chain

This study analyzed DNA minicircles of Mexican isolates ofL. (Leishmania) mexicanato look for genetic differences between strains isolated from patients with diffuse cutaneous (DCL) and localized (LCL) leishmaniasis. The kDNA was analyzed using polymerase chain reaction (PCR), restriction fragment polymorphism analysis of the PCR products (PCR-RFLP) and the PCR products were sequenced. In the PCR with primers specific for the subgenusLeishmania, the Mexican isolates gave higher amplification products than the otherL. mexicanacomplex strains and with specific primers for theL. mexicanacomplex they were poorly amplified. In the PCR-RFLP analysis with theEco RV,Hae III, andMbo Iendonucleases, the Mexican isolates displayed similar restriction patterns, but different from the patterns of the other members of theL. mexicanacomplex. In the phylogenetic tree constructed, the kDNA sequences of the Mexican clones formed two groups including sequences of LCD or LCL clones, apart from the otherL. mexicanacomplex members. These results suggest that the kDNA minicircles of the Mexican isolates are more polymorphic than the kDNA of other members of theL. mexicanacomplex and have different recognition sites for the restriction enzymes used in this study.

scholarly journals First Report of a Group 16SrII Phytoplasma Infecting Chickpea in Oman

Plant Disease ◽  
2006 ◽  
Vol 90 (7) ◽  
pp. 973-973 ◽  
Author(s):  
N. A. Al-Saady ◽  
A. M. Al-Subhi ◽  
A. Al-Nabhani ◽  
A. J. Khan
Keyword(s):  
Nested Pcr ◽  
Animal Feed ◽  
Restriction Enzymes ◽  
Universal Primers ◽  
Polymorphism Analysis ◽  
Disease Symptoms ◽  
First Report ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Group 2

Chickpea (Cicer arietinum), locally known as “Dungo”, is grown for legume and animal feed mainly in the interior region of Oman. During February 2006, survey samples of chickpea leaves from plants showing yellows disease symptoms that included phyllody and little leaf were collected from the Nizwa Region (175 km south of Muscat). Total nucleic acid was extracted from asymptomatic and symptomatic chickpea leaves using a cetyltrimethylammoniumbromide method with modifications (3). All leaf samples from eight symptomatic plants consistently tested positive using a polymerase chain reaction assay (PCR) with phytoplasma universal primers (P1/P7) that amplify a 1.8-kb phytoplasma rDNA product and followed by nested PCR with R16F2n/R16R2 primers yielding a product of 1.2 kb (2). No PCR products were evident when DNA extracted from healthy plants was used as template. Restriction fragment length polymorphism analysis of nested PCR products by separate digestion with Tru9I, HaeIII, HpaII, AluI, TaqI, HhaI, and RsaI restriction enzymes revealed that a phytoplasma belonging to group 16SrII peanut witches'-broom group (2) was associated with chickpea phyllody and little leaf disease in Oman. Restriction profiles of chickpea phytoplasma were identical with those of alfalfa witches'-broom phytoplasma, a known subgroup 16SrII-B strain (3). To our knowledge, this is the first report of phytoplasma infecting chickpea crops in Oman. References: (1) A. J. Khan et al. Phytopathology, 92:1038, 2002. (2). I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998 (3) M. A. Saghai-Maroof et al. Proc. Natl. Acad. Sci. USA. 81:8014, 1984.

scholarly journals Detection of Phytoplasmas in Declining Pears in Southern Australia

Plant Disease ◽  
1997 ◽  
Vol 81 (3) ◽  
pp. 254-258 ◽  
Author(s):  
B. Schneider ◽  
K. S. Gibb
Keyword(s):  
Nested Pcr ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Pcr Analysis ◽  
Pcr Assay ◽  
Specific Primers ◽  
Pear Tree ◽  
Pear Trees ◽  
Polymerase Chain

Forty-nine pear tree samples collected in Victoria, most of them showing decline symptoms, were tested by polymerase chain reaction (PCR) analysis to detect phytoplasmas. Two universal phytoplasma-specific primer pairs, fP1/rP7 and fU5/rU3, were tested, but only fU5/rU3 amplified the phytoplasma DNA adequately. Nested PCR with universal and group-specific primers, however, proved more effective. Thirty pear trees reacted positively in a nested PCR assay. Restriction fragment length polymorphism (RFLP) analysis with the restriction enzymes MseI and AluI of the PCR fragment amplified with the primer pair fU5/rU3 revealed patterns identical to those from the sweet potato little leaf phytoplasma. This is the first report of a phytoplasma in pear in Australia.

scholarly journals Restriction fragment length polymorphism analysis of isolates of infectious bursal disease viruses from Turkey

2012 ◽  
Vol 48 (No. 12) ◽  
pp. 359-362 ◽  
Author(s):  
G. Ozbes ◽  
Ertas HB ◽  
A. Muzo
Keyword(s):  
Rflp Analysis ◽  
Infectious Bursal Disease ◽  
Polymorphism Analysis ◽  
Rt Pcr ◽  
Vp2 Gene ◽  
Infectious Bursal Disease Viruses ◽  
Pcr Rflp ◽  
Bursal Disease ◽  
Polymerase Chain ◽  
Rflp Assay

Infectious bursal disease Virus (IBDV) specific reverse transcriptase/polymerase chain recation (RT/PCR) positive 40 broiler bursa fabricius samples obtained from a commercially reared flock were investigated for genetic diversity by PCR-RFLP assay. The assay amplifies a 743 bp fragment of the IBDV VP2 gene. The RFLP profiles of 40 of these positive samples were determined using the enzyme MboI. Most of the viruses had the same RFLP with the MboI enzyme. RFLP analysis of the isolates produced two different band profiles. The results of this study showed that little genetic heterogeneity exists among IBDV strains in a infected flock.

scholarly journals Survey of mutations in prolificacy genes in Santa Ines and Morada Nova sheep

2017 ◽  
Vol 69 (4) ◽  
pp. 1047-1053
Author(s):  
G.M.L. Holanda ◽  
J.C. Oliveira ◽  
D.M.F. Silva ◽  
S.S.N. Rocha ◽  
V. Pandolfi ◽  
...  
Keyword(s):  
Restriction Site ◽  
Fragment Length Polymorphism ◽  
Nucleotide Sequencing ◽  
Chain Reaction ◽  
Specific Primers ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Santa Inês ◽  
Restriction Fragment Length

ABSTRACT Polymorphisms in the BMP-15 gene related to Galway (FecXG) and Inverdale (FecXI) and in the BMPR-1B gene known as Booroola (FecB) mutations were investigated using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) method, on sheep from the breeds Santa Inês (n= 574) and Morada Nova (n=282). DNA was extracted and amplified through PCR with specific primers that introduced a restriction site in association with the mutation. The PCR products were submitted to endonucleases. The experiment found no FecXG and FecXI mutations. Six samples of animals with multiple offspring/birth history presented polymorphism for FecB similar to control samples, but this pattern was not confirmed by nucleotide sequencing. Although the absence of these mutations in the studied breeds, other factors related to prolificacy should be investigated to explain the inherent prolificity mechanisms.

Genetic polymorphisms of cytochrome P450 enzymes: CYP2C9, CYP2C19, CYP2D6, CYP3A4, and CYP3A5 in the Croatian population

2017 ◽  
Vol 32 (1) ◽  
Author(s):  
Lana Ganoci ◽  
Tamara Božina ◽  
Nikica Mirošević Skvrce ◽  
Mila Lovrić ◽  
Petar Mas ◽  
...  
Keyword(s):  
Rflp Analysis ◽  
The Other ◽  
Published Data ◽  
Cytochrome P450 Enzymes ◽  
Chain Reaction ◽  
Allelic Variants ◽  
Mediterranean Populations ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
European Populations

AbstractBackground:Data on the frequency of pharmacogenetic polymorphisms in the Croatian population are limited. We determined and analyzed frequencies for the most importantMethods:2637 subjects were included. Genotyping was performed by real-time polymerase chain reaction (PCR) using TaqMan® DME or TaqMan® SNP Genotyping Assays, and by PCR, and PCR-RFLP analysis.Results:ForConclusions:The frequency of the CYP allelic variants, genotypes, and predicted phenotypes in the Croatian population is in accordance with the other European populations, between the values of published data for Middle European and Mediterranean populations.

scholarly journals The Application of Polymerase Chain Reaction – Restriction Fragment Polymorphisms (PCR-RFLP) to Determine Genetic Diversity of Madura Cattle in Sapudi Island

2015 ◽  
Vol 18 (1) ◽  
pp. 70
Author(s):  
Tety Hartatik ◽  
Slamet Diah Volkandari ◽  
S. Sumadi ◽  
W. Widodo
Keyword(s):  
Genetic Diversity ◽  
Polymerase Chain Reaction ◽  
Restriction Fragment ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Cytb Gene ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Haplotype A

The aim of this study was to determine genetic diversity of Madura cattle using Polymerase Chain Reaction – Restriction Fragment Length Polymorphisms (PCR-RFLP) analysis of the cytochrome b (cytb) gene. Samples used for the experiments were blood of 43 cattle that consist of 15 cattle obtained from Madura Island, 23 cattle from Sapudi Island, and 5 Limousin-Madura (Limura) cattle. A fragment of 464 base pair of cytb gene was amplifi ed by forward primer L14735 and reverse primer H15149. The PCR product was digested with TaqIand HinfI restriction enzymes to identify genetic patterns. Data of PCR-RFLP showed two haplotypes, that were A and B, in cattle obtained from both Madura Island and Sapudi Island. The frequencies of haplotype A and B of cattle from Sapudi Island were 69.57% and 30.47%, respectively. More diverse frequencies were observed in cattle obtained from Madura Island, where haplotype A and B were 86.67% and 13.33%, respectively. In this experiment, Limura cattle had only haplotype A. As a conclusion, PCR-RFLP of the cytb gene had been able to determine a genetic diversity of Madura cattle. Key words: Genetic diversity, Madura cattle, haplotype.

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 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.

scholarly journals Flagellin gene polymorphism analysis ofCampylobacter jejuniinfecting man and other hosts and comparison with biotyping and somatic antigen serotyping

1994 ◽  
Vol 113 (2) ◽  
pp. 221-234 ◽  
Author(s):  
R. J. Owen ◽  
C. Fitzgerald ◽  
K. Sutherland ◽  
P. Borman
Keyword(s):  
Polymorphism Analysis ◽  
Flagellin Gene ◽  
Novel Approach ◽  
Length Polymorphisms ◽  
Pcr Rflp ◽  
Pcr Products ◽  
Enteric Infections ◽  
Polymerase Chain ◽  
Somatic Antigens ◽  
Dna Pcr

SUMMARYFlagellin gene sequence polymorphisms were used to discriminate amongst 77 strains ofCampylobacter jejunifrom sporadic and outbreak-associated human enteric infections, and from chickens, sheep and calves. The results were assessed in relation to Lior biotyping and serotyping (Penner somatic antigens). Eight DNA PCR-RFLP patterns (genotypes) were identified by analysis ofHinfI fragment length polymorphisms in flagellin gene (flaA) polymerase chain reaction (PCR) products. One genotype (F-l) was a feature of 55% of strains. Strains within the genotypes were heterogeneous with respect to somatic antigens with 12 serogroups represented amongst theC. jejuniisolates offlaA type F-l. Serogroups Pen 1. 2 and 23 were the commonest (45%) amongst the 20 different serogroups represented. Several unique clusters of isolates with diverse biotypes were defined, and one cluster (F-7/Pen 23) contained epidemiologically implicated outbreak strains as well as sheep and calf isolates. We conclude thatHinfIflaA typing is reproducible and offers high typability, and its combination with serogrouping provides a novel approach to characterizing isolates ofC. jejuniwith improved discrimination.

Rapid and reliable identification of rice genomes by RFLP analysis of PCR-amplified Adh genes

Genome ◽  
2001 ◽  
Vol 44 (6) ◽  
pp. 1136-1142 ◽  
Author(s):  
Song Ge ◽  
Tao Sang ◽  
Bao-rong Lu ◽  
De-yuan Hong
Keyword(s):  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Pcr Primers ◽  
Specific Pcr ◽  
Adh Genes ◽  
Pcr Rflp ◽  
Restriction Sites ◽  
Pcr Products ◽  
Adh Gene ◽  
Genus Oryza

The rice genus (Oryza L.) consists of 24 species with 10 recognized genome types. With the realization of many useful genes in species of wild rice, continuous efforts have been made to understand their genomic composition and relationships. However, the identification of rice genomes has often been difficult owing to complex morphological variation and formation of allotetraploids. Here we propose a rapid and reliable method for identifying rice genomes based on the restriction sites of PCR-amplified Adh genes. The experimental procedure was as follows: (i) amplify a portion of Adh1 and Adh2 genes with the locus-specific PCR primers; (ii) digest PCR products with restriction enzymes that distinguish different genomes; and (iii) run the digested products on 1.4% agarose gel, and photograph. Using various combinations of restriction digestion of the two Adh genes, all of the rice genomes can be identified.Key words: Adh gene, genome, identification, Oryza L., PCR–RFLP.

Sign in / Sign up

Export Citation Format

Share Document