Parameters affecting the yield of DNA from human blood

Gustafson, Steven Wayne; Proper, Jacqueline A.; Bowie, E. J. W.; Sommer, S. S.

doi:10.1016/0003-2697(87)90272-7

Cited by 163 publications

(80 citation statements)

References 9 publications

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“…DNA from blood of fifty unrelated AfricanAmerican individuals (AA1-AA50) were extracted with phenol chloroform (Gustafson et al, 1987). The work was approved by the City of Hope's Human Subjects Use Committee.…”

Section: Methodsmentioning

confidence: 99%

Identification of Novel Variants of the Flavin-Containing Monooxygenase Gene Family in African Americans

Furnes¹,

Feng²,

Sommer³

et al. 2003

Drug Metab Dispos

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ABSTRACT:Sequence polymorphisms in enzymes involved in drug metabolism have been widely implicated in the differences observed in the sensitivity to various xenobiotics. The flavin-containing monooxygenase (FMO) gene family in humans catalyzes the monooxygenation of numerous N-, P-and S-containing drugs, pesticides, and environmental toxicants. Six genes (FMO1-6) have been identified so far, but the major alleles of FMO2 and FMO6 encode nonfunctional proteins due to a nonsense mutation and splice-site abnormalities, respectively. Data on structural variants exist for human FMO2 and 3, whereas very little is known about the other FMO genes. FMO1-6 were scanned in 50 individuals of African-American descent using the method, detection of virtually all mutationssingle-strand conformational polymorphism. A total of 49 sequence variants were identified in a total 1.35 megabases of scanned sequence, of which 29 were variants affecting protein structure or expression. Some of these are expected to affect the activity of the protein, including a nonsense mutation in FMO1 (R502X) and missense mutations in FMO1 (I303T), FMO4 (E339Q), and FMO5 (P457L) that occur in highly conserved amino acids. Additional deleterious substitutions in FMO2 (del337G) and FMO6 (Q105X) were also identified. Multiple structural variants in the FMO gene family were observed in this African-American sample. Some of the substitutions identified in this study might be useful markers in future association studies assessing sensitivity to environmental toxicants and common disease.

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Section: Methodsmentioning

confidence: 99%

Identification of Novel Variants of the Flavin-Containing Monooxygenase Gene Family in African Americans

Furnes¹,

Feng²,

Sommer³

et al. 2003

Drug Metab Dispos

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“…Primers were designed for PCR amplification of the 11 exons and part of the flanking intron sequences of the human ACAD8 gene (primer sequences are available on request brage@biobase.dk). Genomic DNA was isolated from cultured fibroblasts or blood samples according to standard methods [16]. PCR reactions were performed under standard conditions in an automated Thermal cycler 480 (Perkin-Elmer, Norwalk, CT) and the PCR products were subjected to direct bi-directional cycle sequencing as described above.…”

Section: Thementioning

confidence: 99%

Identification of isobutyryl-CoA dehydrogenase and its deficiency in humans

Nguyen

Andresen

Corydon

et al. 2002

Molecular Genetics and Metabolism

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The acyl-CoA dehydrogenases (ACDs) are a family of related enzymes that catalyze the a,b-dehydrogenation of acyl-CoA esters. Two homologues active in branched chain amino acid metabolism have previously been identified. We have used expression in Escherichia coli to produce a previously uncharacterized ACD-like sequence (ACAD8) and define its substrate specificity. Purified recombinant enzyme had a k cat =K m of 0.8, 0.23, and 0.04 (lM À1 s À1 ) with isobutyryl-CoA, (S) 2-methylbutyryl-CoA, and n-propionylCoA, respectively, as substrates. Thus, this enzyme is an isobutyryl-CoA dehydrogenase. A single patient has previously been described whose fibroblasts exhibit a specific deficit in the oxidation of valine. Amplified ACAD8 cDNA made from patient fibroblast mRNA was homozygous for a single nucleotide change (905G > A) in the ACAD8 coding region compared to the sequence from control cells. This encodes an Arg302Gln substitution in the full-length protein (position 280 in the mature protein), a position predicted by molecular modeling to be important in subunit interactions. The mutant enzyme was stable but inactive when expressed in E. coli. It was also stable and appropriately targeted to mitochondria, but inactive when expressed in mammalian cells. These data confirm further the presence of a separated ACD in humans specific to valine catabolism (isobutyryl-CoA dehydrogenase, IBDH), along with the first enzymatic and molecular confirmation of a deficiency of this enzyme in a patient. Ó

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“…Blood from the cancer patients and the remaining controls was obtained frozen in heparin or EDTA-containing tubes. After thawing, the samples were subjected to lysis and proteinase digestion before extraction with phenol and chloroform [12]. DNA was isolated after ethanol precipitation.…”

Section: Introductionmentioning

confidence: 99%

Genetic polymorphism of cytochrome P4502E1 in a Swedish population

et al. 1993

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Genetic polymorphism of CYPZEI was investigated among 195 Swedish patients with lung cancer and 206 controls. Three different polymorphic sites were found, all in introns, using RFLP and the restriction enzymes DruI, RraI and TuqI. The frequencies of the rare alleles were 0.08-0.18 and much lower than previously described among Japanese. No significant difference in distribution of the polymorphic alleles between controls and lung cancer patients was evident, in contrast to results of a previous Japanese study. However, examination of a polymorphic site in the 5'-flanking region, within a putative binding motif for the hepatic transcription factor HNF-1, revealed a significantly less frequent distribution of the mutated allele (cz) among the lung cancer patients as compared to controls. It is concluded that major interethnic differences exist in the genetic polymorphism of CYPZEI and that people carrying the ct allele might be at lower risk for developing lung cancer.

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Parameters affecting the yield of DNA from human blood

Cited by 163 publications

References 9 publications

Identification of Novel Variants of the Flavin-Containing Monooxygenase Gene Family in African Americans

Identification of Novel Variants of the Flavin-Containing Monooxygenase Gene Family in African Americans

Identification of isobutyryl-CoA dehydrogenase and its deficiency in humans

Genetic polymorphism of cytochrome P4502E1 in a Swedish population

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