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Long-range PCR facilitates the identification of PMS2-specific mutations

Clendenning, M; Hampel, H; LaJeunesse, J; Lindblom, A; Lockman, J; Nilbert, Mef LU ; Senter, L; Sotamaa, K and de la Chapelle, A (2006) In Human Mutation 27(5). p.490-495
Abstract
Mutations within the DNA mismatch repair gene, "postmeiotic segregation increased 2" (PMS2), have been associated with a predisposition to hereditary nonpolyposis colorectal cancer (HNPCC; Lynch syndrome). The presence of a large family of highly homologous PMS2 pseudogenes has made previous attempts to sequence PMS2 very difficult. Here, we describe a novel method that utilizes long-range PCR as a way to preferentially amplify PMS2 and not the pseudogenes. A second, exon-specific, amplification from diluted long-range products enables us to obtain a clean sequence that shows no evidence of pseudogene contamination. This method has been used to screen a cohort of patients whose tumors were negative for the PMS2 protein by... (More)
Mutations within the DNA mismatch repair gene, "postmeiotic segregation increased 2" (PMS2), have been associated with a predisposition to hereditary nonpolyposis colorectal cancer (HNPCC; Lynch syndrome). The presence of a large family of highly homologous PMS2 pseudogenes has made previous attempts to sequence PMS2 very difficult. Here, we describe a novel method that utilizes long-range PCR as a way to preferentially amplify PMS2 and not the pseudogenes. A second, exon-specific, amplification from diluted long-range products enables us to obtain a clean sequence that shows no evidence of pseudogene contamination. This method has been used to screen a cohort of patients whose tumors were negative for the PMS2 protein by immunohistochemistry and had not shown any mutations within the MLH1 gene. Sequencing of the PMS2 gene from 30 colorectal and I I endometrial cancer patients identified 10 novel sequence changes as well as 17 sequence changes that had previously been identified. In total, putative pathologic mutations were detected in 11 of the 41 families. Among these were five novel mutations, c.705+1G > T, c.736-741del6ins11, c.862_863del, c.1688G > T, and c.2007-IG > A. We conclude that PMS2 mutation detection in selected Lynch syndrome and Lynch syndrome-like patients is both feasible and desirable. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
polymorphisms, pseudogenes, PMS2, Lynch syndrome
in
Human Mutation
volume
27
issue
5
pages
490 - 495
publisher
John Wiley & Sons
external identifiers
  • wos:000237275900014
  • pmid:16619239
  • scopus:33646372203
ISSN
1059-7794
DOI
10.1002/humu.20318
language
English
LU publication?
yes
id
7d045c59-d67a-4e9f-877e-eb27d2a9da7f (old id 410462)
date added to LUP
2007-10-01 19:17:35
date last changed
2019-05-14 01:52:35
@article{7d045c59-d67a-4e9f-877e-eb27d2a9da7f,
  abstract     = {Mutations within the DNA mismatch repair gene, "postmeiotic segregation increased 2" (PMS2), have been associated with a predisposition to hereditary nonpolyposis colorectal cancer (HNPCC; Lynch syndrome). The presence of a large family of highly homologous PMS2 pseudogenes has made previous attempts to sequence PMS2 very difficult. Here, we describe a novel method that utilizes long-range PCR as a way to preferentially amplify PMS2 and not the pseudogenes. A second, exon-specific, amplification from diluted long-range products enables us to obtain a clean sequence that shows no evidence of pseudogene contamination. This method has been used to screen a cohort of patients whose tumors were negative for the PMS2 protein by immunohistochemistry and had not shown any mutations within the MLH1 gene. Sequencing of the PMS2 gene from 30 colorectal and I I endometrial cancer patients identified 10 novel sequence changes as well as 17 sequence changes that had previously been identified. In total, putative pathologic mutations were detected in 11 of the 41 families. Among these were five novel mutations, c.705+1G > T, c.736-741del6ins11, c.862_863del, c.1688G > T, and c.2007-IG > A. We conclude that PMS2 mutation detection in selected Lynch syndrome and Lynch syndrome-like patients is both feasible and desirable.},
  author       = {Clendenning, M and Hampel, H and LaJeunesse, J and Lindblom, A and Lockman, J and Nilbert, Mef and Senter, L and Sotamaa, K and de la Chapelle, A},
  issn         = {1059-7794},
  keyword      = {polymorphisms,pseudogenes,PMS2,Lynch syndrome},
  language     = {eng},
  number       = {5},
  pages        = {490--495},
  publisher    = {John Wiley & Sons},
  series       = {Human Mutation},
  title        = {Long-range PCR facilitates the identification of PMS2-specific mutations},
  url          = {http://dx.doi.org/10.1002/humu.20318},
  volume       = {27},
  year         = {2006},
}