Long-range PCR facilitates the identification of PMS2-specific mutations
(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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/410462
- author
- Clendenning, M ; Hampel, H ; LaJeunesse, J ; Lindblom, A ; Lockman, J ; Nilbert, Mef LU ; Senter, L ; Sotamaa, K and de la Chapelle, A
- organization
- publishing date
- 2006
- 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 Inc.
- 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
- 2016-04-01 11:42:24
- date last changed
- 2022-01-26 17:00:47
@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}},
keywords = {{polymorphisms; pseudogenes; PMS2; Lynch syndrome}},
language = {{eng}},
number = {{5}},
pages = {{490--495}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Human Mutation}},
title = {{Long-range PCR facilitates the identification of PMS2-specific mutations}},
url = {{http://dx.doi.org/10.1002/humu.20318}},
doi = {{10.1002/humu.20318}},
volume = {{27}},
year = {{2006}},
}