Research article summary (published 30 Oct 1999):

Application of an improved cDNA competition technique to identify prostate cancer-associated gene.

Full Abstract

A technique to improve cDNA library screening was developed by using mixed probes derived from two closely related cDNA populations of high-metastatic MAT-LyLu and low-metastatic AT-1 Dunning R3227 rat prostate cancer sublines. The technique required the generation of a cDNA library from each subline followed by polymerase chain reaction (PCR) amplification of the cDNA insert population. The PCR products derived from the first library were radiolabeled and mixed with an excess amount of PCR products from the second library. The mixture and an excess amount of both the lambda and pBluescript DNA were used as a probe to screen the first cDNA library. This mixed probe (designated the competition probe) differentially cross-hybridized with the plaque lift of the screened first cDNA library. Weak radioactive signals indicated the cross-hybridization of cDNA sequences common to the competition probe mixture and the first cDNA library, whereas strong signals implied unhybridized unique or abundant cDNA sequences in the first cDNA library. The reproducibility of this technique was confirmed by showing that the full-length cDNA clones were associated with the phenotype of the screened first cell line. The isolated clones were characterized as rat nucleolar protein, rat mitochondrial genes coding for 16S and 12S rRNAs, and rat tRNAs specific for valine and phenyl-alanine. This result is consistent with the fact that the first cell line, MAT-LyLu, is metabolically more active than are AT-1 cells because of higher gene dosage or amplification of nucleolar and mitochondrial RNA and its associated genes. Another clone which had a strong signal represented a novel gene associated with the MAT-LyLu cancer phenotype.

Author information

Author/s: Rinaldy, A R (AR); Steiner, M S (MS);

Affiliation: University of Tennessee Urologic Research Laboratories, College of Medicine, University of Tennessee, Memphis 38163, USA. ARinaldy(-atsign-)UTMEM1.UTMEM.EDU

Grants: CA 43769 (Agency:NCI NIH HHS)

Journal and publication information

Publication Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.

Journal: DNA and cell biology (DNA Cell Biol), published in UNITED STATES. (Language: eng)

Reference: 1999-Nov; vol 18 (issue 11) : pp 829-36

Dates: Created 2000/01/04; Completed 2000/01/04; Revised 2007/11/14;

PMID: 10595396, status: MEDLINE (last retrieval date: 2/18/2009, IMS Date: )

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Animals Base Sequence DNA, Complementary - genetics DNA, Mitochondrial - genetics DNA, Neoplasm - genetics Gene Library Male Molecular Probe Techniques Molecular Sequence Data Nuclear Proteins - genetics

Nucleic Acid Hybridization - methods Polymerase Chain Reaction - methods Prostatic Neoplasms - chemistry; genetics RNA, Neoplasm - analysis; isolation & purification RNA, Ribosomal - genetics RNA, Ribosomal, 16S - genetics RNA, Transfer, Phe - genetics RNA, Transfer, Val - genetics Rats Tumor Cells, Cultured

Associated Chemicals: DNA, Complementary (0) ; DNA, Mitochondrial (0) ; DNA, Neoplasm (0) ; Nuclear Proteins (0) ; RNA, Neoplasm (0) ; RNA, Ribosomal (0) ; RNA, Ribosomal, 16S (0) ; RNA, Transfer, Phe (0) ; RNA, Transfer, Val (0) ; RNA, ribosomal, 12S (0)

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