PCR Protocol for PfCRT Pyrosequencing
Date
2012-02-09Publisher
University of Maryland School of Medicine. Center for Vaccine Development. Malaria GroupType
OtherMetadata
Show full item recordOther Titles
SUBTITLE: (codons 72, 74, 75, 76, 220,271,326, and 371)Description
"Provides a method for performing PCR on genomic DNA from dried blood spots on Whatman 3MM filter paper ... for subsequent genotyping of pfcrt codons ..."Includes the following sections: 1.0 Purpose; 2.0 References; 3.0 Materials; 4.0 Equipment; 5.0 Safety and Precautions; 6.0 Procedure; 6.1 DNA extraction from blood spotted filter papers; 6.2 Setting up PCR Reactions
Keyword
pfcrt codon 72pfcrt codon 74
pfcrt codon 75
pfcrt codon 76
pfcrt codon 220
pfcrt codon 271
pfcrt codon 326
pfcrt codon 356
pfcrt codon 371
pyrosequencingUniversity of Maryland, Baltimore. Center for Vaccine Development (CVD)Genotyping Techniques--methodsPlasmodium falciparum--geneticsPolymerase Chain ReactionSequence Analysis--methods
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http://hdl.handle.net/10713/7042Collections
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PCR and allele-specific restriction analysis (ASRA) of pfcrtcodons 220 and 271University of Maryland School of Medicine. Center for Vaccine Development. Malaria Group, 2012-02-08
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A precedented nuclear genetic code with all three termination codons reassigned as sense codons in the syndinean Amoebophrya sp. Ex Karlodinium veneficumAmoebophrya is part of an enigmatic, diverse, and ubiquitous marine alveolate lineage known almost entirely from anonymous environmental sequencing. Two cultured Amoebophrya strains grown on core dinoflagellate hosts were used for transcriptome sequencing. BLASTx using different genetic codes suggests that Amoebophyra sp. ex Karlodinium veneficum uses the three typical stop codons (UAA, UAG, and UGA) to encode amino acids. When UAA and UAG are translated as glutamine about half of the alignments have better BLASTx scores, and when UGA is translated as tryptophan one fifth have better scores. However, the sole stop codon appears to be UGA based on conserved genes, suggesting contingent translation of UGA. Neither host sequences, nor sequences from the second strain, Amoebophrya sp. ex Akashiwo sanguinea had similar results in BLASTx searches. A genome survey of Amoebophyra sp. ex K. veneficum showed no evidence for transcript editing aside from mitochondrial transcripts. The dynein heavy chain (DHC) gene family was surveyed and of 14 transcripts only two did not use UAA, UAG, or UGA in a coding context. Overall the transcriptome displayed strong bias for A or U in third codon positions, while the tRNA genome survey showed bias against codons ending in U, particularly for amino acids with two codons ending in either C or U. Together these clues suggest contingent translation mechanisms in Amoebophyra sp. ex K. veneficum and a phylogenetically distinct instance of genetic code modification. © 2019 Tsvetan R. Bachvaroff. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
