Expression of Human Intestinal mRNA Transcripts during Development: Analysis by a Semiquantitative RNA Polymerase Chain Reaction Method

Wang, Yangxi; Harvey, Clare B.; Rousset, Monique; Swallow, Dallas M.

doi:10.1203/00006450-199410000-00018

Cited by 44 publications

(27 citation statements)

References 34 publications

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“…The level of MCM6 transcript in the small intestine of children was comparable to that of the adults, both children and adults showing a lower level of expression than the fetuses. This pattern of expression contrasts with that of lactase which shows a very restricted distribution, occurring at detectable levels only in the intestine, and at high levels only in the small intestine of children and lactase persistent adults [15,17] (Fig. 4).…”

Section: Mcm6 Expression In Human Tissuesmentioning

confidence: 78%

“…Semiquantitative RT-PCR was performed using the method described previously [15] except that the RT was performed at 37°C using 1 Ixg total RNA as starting material for 10 PCR reactions and, for MCM6 only, 30 rounds of amplification (which gave a linear dose response for this transcript). Primer sequences for dipeptidylpeptidase IV (DPP4) and lactase-phlorizin hydrolase (LCT) were as described previously.…”

Section: Rna Samples and Rt-pcr Analysismentioning

confidence: 99%

“…The human tissues were as described previously [1,[15][16][17]. Semiquantitative RT-PCR was performed using the method described previously [15] except that the RT was performed at 37°C using 1 Ixg total RNA as starting material for 10 PCR reactions and, for MCM6 only, 30 rounds of amplification (which gave a linear dose response for this transcript).…”

Section: Rna Samples and Rt-pcr Analysismentioning

confidence: 99%

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Characterisation of a human homologue of a yeast cell division cycle gene, MCM6, located adjacent to the 5′ end of the lactase gene on chromosome 2q21

Harvey¹,

Wang²,

Darmoul

et al. 1996

FEBS Letters

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Section: Mcm6 Expression In Human Tissuesmentioning

confidence: 78%

Section: Rna Samples and Rt-pcr Analysismentioning

confidence: 99%

Section: Rna Samples and Rt-pcr Analysismentioning

confidence: 99%

See 1 more Smart Citation

Characterisation of a human homologue of a yeast cell division cycle gene, MCM6, located adjacent to the 5′ end of the lactase gene on chromosome 2q21

Harvey¹,

Wang²,

Darmoul

et al. 1996

FEBS Letters

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“…The level of sucrase-isomaltase and lactase mRNA present in the samples was determined using the semiquantitative RNA-PCR method which is described in detail elsehwere. 22 Briefly, total RNA was prepared from a frozen portion of each biopsy specimen (approximately 5 mg) using a guanadinium thiocyanate-phenol-chloroform method. 23 Reverse transcription of 2 ,ug RNA was primed using random primers and cDNA corresponding to 0 5 ,ug RNA was used for PCR.…”

Section: Sds-polyacrylamide Gel Electrophoresis (Page)mentioning

confidence: 99%

Studies on the expression of intestinal lactase in different individuals.

Harvey

Wang

Hughes

et al. 1995

Gut

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Sixty one duodenal biopsy specimens were examined for the expression of lactase at the level of enzyme activity, protein, and messenger RNA. Of the 51 samples with normal villous architecture, 39 were lactase persistent, 11 were nonpersistent (adult type hypolactasia), and one was of indeterminate status. All the lactase persistent individuals showed high mRNA and a high level of the lactase protein as detected by sodium dodecyl sulphate polyacrylamide gel electrophoresis. All the 11 non-persistent individuals tested showed a low level of lactase protein.Nine of the 10 samples tested showed low mRNA and one high mRNA. These results suggest that the lactase persistence polymorphism is controlled at the level of the expression of the lactase gene, though there may be some heterogeneity of the lactase non-persistence phenotype. (Gut 1995; 36: 28-33)

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“…Relative mRNA transcript levels were determined using PCR products obtained by RT-PCR performed under the general conditions described previously (Wang et al 1994) with appropriate controls and 'no RT' blanks. The primers for the RT-PCR are located in exons 1 and 3 of LCT, and 30 cycles of amplification was selected to produce sufficient template DNA for sequencing.…”

Section: Determination Of Allelic Expressionmentioning

confidence: 99%

The Causal Element for the Lactase Persistence/ non‐persistence Polymorphism is Located in a 1 Mb Region of Linkage Disequilibrium in Europeans

Poulter

Hollox

Harvey

et al. 2003

Annals of Human Genetics

112

121

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SummaryExpression of lactase in the intestine persists into adult life in some people and not others, and this is due to a cisacting regulatory polymorphism. Previous data indicated that a mutation leading to lactase persistence had occurred on the background of a 60 kb 11-site LCT haplotype known as A (Hollox et al. 2001). Recent studies reported a 100% correlation of lactase persistence with the presence of the T allele at a CT SNP at − 14 kb from LCT, in individuals of Finnish origin, suggesting that this SNP may be causal of the lactase persistence polymorphism, and also reported a very tight association with a second SNP (GA -22 kb) (Enattah et al. 2002). Here we report the existence of a one megabase stretch of linkage disequilibrium in the region of LCT and show that the -14 kb T allele and the -22 kb A allele both occur on the background of a very extended A haplotype. In a series of Finnish individuals we found a strong correlation (40/41 people) with lactose digestion and the presence of the T allele. The T allele was present in all 36 lactase persistent individuals from the UK (phenotyped by enzyme assay) studied, 31/36 of whom were of Northern European ancestry, but not in 11 non-persistent individuals who were mainly of non-UK ancestry. However, the CT heterozygotes did not show intermediate lactase enzyme activity, unlike those previously phenotyped by determining allelic transcript expression. Furthermore the one lactase persistent homozygote identified by having equally high expression of A and B haplotype transcripts, was heterozygous for CT at the − 14 kb site. SNP analysis across the 1 megabase region in this person showed no evidence of recombination on either chromosome between the -14 kb SNP and LCT. The combined data shows that although the -14 kb CT SNP is an excellent candidate for the cause of the lactase persistence polymorphism, linkage disequilibrium extends far beyond the region searched so far. In addition, the CT SNP does not, on its own, explain all the variation in expression of LCT, suggesting the possibility of genetic heterogeneity. *

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Expression of Human Intestinal mRNA Transcripts during Development: Analysis by a Semiquantitative RNA Polymerase Chain Reaction Method

Cited by 44 publications

References 34 publications

Characterisation of a human homologue of a yeast cell division cycle gene, MCM6, located adjacent to the 5′ end of the lactase gene on chromosome 2q21

Characterisation of a human homologue of a yeast cell division cycle gene, MCM6, located adjacent to the 5′ end of the lactase gene on chromosome 2q21

Studies on the expression of intestinal lactase in different individuals.

The Causal Element for the Lactase Persistence/ non‐persistence Polymorphism is Located in a 1 Mb Region of Linkage Disequilibrium in Europeans

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