Multiplex polymerase chain reaction-based deletion analysis of spontaneous, gamma ray- and alpha-induced hprt mutants of CHO-K1 cells

Jeffrey L. Schwartz, Jacob Rotmensch, Juan Sun, Jie An, Zhidong Xu, Yongjia Yu, Abraham W. Hsie

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Independent Chinese hamster ovary (CHO)-K1 cell mutants at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus were isolated from untreated, 60Co γ ray-and 212Bi α-exposed cells and the genetic changes underlying the mutation determined by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. In the 71 spontaneous mutants analyzed, 77.5% of the clones showed no change in exon number or size, 15.5% showed a loss of a single exon, 4.2% showed a loss of 2-8 exons, and 2.8% showed loss of all nine hprt exons (total gene deletion). Exposure to 6 Gy of γ rays, which reduced survival levels to 10%, produced a significantly different deletion spectrum that was shifted toward deletions with 45% of the 20 mutants analyzed showing a loss of a single exon and 30% showing a loss of all nine exons. Exposure to 2 Gy α radiation from 212Bi, a 220Rn daughter, a dose which also reduced survival levels to about 10%, resulted in a deletion spectrum similar to the γ-ray spectrum in that more than 75% of the 49 mutants analyzed were deletions. The α spectrum, however, was significantly different from both the spontaneous and γ spectra with 55.1% of the α mutants showing a loss of all nine exons, 10.2% showing loss of a single exon, and 14.3% showing loss of 2-8 exons. Thus, α-radiation appears to produce larger intragenic deletions than γ radiation. The results suggest that intragenic deletion size should be considered when low- and high linear energy transfer (LET) mutation spectra are compared.

Original languageEnglish (US)
Pages (from-to)537-540
Number of pages4
JournalMutagenesis
Volume9
Issue number6
DOIs
StatePublished - Nov 1994

Fingerprint

Hypoxanthine Phosphoribosyltransferase
Ovary
Polymerase Chain Reaction
Multiplex Polymerase Chain Reaction
Polymerase chain reaction
Gamma Rays
Cricetulus
Mutant
Gamma rays
Deletion
Exons
Cell
Half line
Radiation
Mutation
Hamster
Energy Transfer
Linear Energy Transfer
Radiation Dosage
Clone

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

Multiplex polymerase chain reaction-based deletion analysis of spontaneous, gamma ray- and alpha-induced hprt mutants of CHO-K1 cells. / Schwartz, Jeffrey L.; Rotmensch, Jacob; Sun, Juan; An, Jie; Xu, Zhidong; Yu, Yongjia; Hsie, Abraham W.

In: Mutagenesis, Vol. 9, No. 6, 11.1994, p. 537-540.

Research output: Contribution to journalArticle

Schwartz, Jeffrey L. ; Rotmensch, Jacob ; Sun, Juan ; An, Jie ; Xu, Zhidong ; Yu, Yongjia ; Hsie, Abraham W. / Multiplex polymerase chain reaction-based deletion analysis of spontaneous, gamma ray- and alpha-induced hprt mutants of CHO-K1 cells. In: Mutagenesis. 1994 ; Vol. 9, No. 6. pp. 537-540.
@article{c2b3519597344710b57233b4cef5093b,
title = "Multiplex polymerase chain reaction-based deletion analysis of spontaneous, gamma ray- and alpha-induced hprt mutants of CHO-K1 cells",
abstract = "Independent Chinese hamster ovary (CHO)-K1 cell mutants at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus were isolated from untreated, 60Co γ ray-and 212Bi α-exposed cells and the genetic changes underlying the mutation determined by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. In the 71 spontaneous mutants analyzed, 77.5{\%} of the clones showed no change in exon number or size, 15.5{\%} showed a loss of a single exon, 4.2{\%} showed a loss of 2-8 exons, and 2.8{\%} showed loss of all nine hprt exons (total gene deletion). Exposure to 6 Gy of γ rays, which reduced survival levels to 10{\%}, produced a significantly different deletion spectrum that was shifted toward deletions with 45{\%} of the 20 mutants analyzed showing a loss of a single exon and 30{\%} showing a loss of all nine exons. Exposure to 2 Gy α radiation from 212Bi, a 220Rn daughter, a dose which also reduced survival levels to about 10{\%}, resulted in a deletion spectrum similar to the γ-ray spectrum in that more than 75{\%} of the 49 mutants analyzed were deletions. The α spectrum, however, was significantly different from both the spontaneous and γ spectra with 55.1{\%} of the α mutants showing a loss of all nine exons, 10.2{\%} showing loss of a single exon, and 14.3{\%} showing loss of 2-8 exons. Thus, α-radiation appears to produce larger intragenic deletions than γ radiation. The results suggest that intragenic deletion size should be considered when low- and high linear energy transfer (LET) mutation spectra are compared.",
author = "Schwartz, {Jeffrey L.} and Jacob Rotmensch and Juan Sun and Jie An and Zhidong Xu and Yongjia Yu and Hsie, {Abraham W.}",
year = "1994",
month = "11",
doi = "10.1093/mutage/9.6.537",
language = "English (US)",
volume = "9",
pages = "537--540",
journal = "Mutagenesis",
issn = "0267-8357",
publisher = "Oxford University Press",
number = "6",

}

TY - JOUR

T1 - Multiplex polymerase chain reaction-based deletion analysis of spontaneous, gamma ray- and alpha-induced hprt mutants of CHO-K1 cells

AU - Schwartz, Jeffrey L.

AU - Rotmensch, Jacob

AU - Sun, Juan

AU - An, Jie

AU - Xu, Zhidong

AU - Yu, Yongjia

AU - Hsie, Abraham W.

PY - 1994/11

Y1 - 1994/11

N2 - Independent Chinese hamster ovary (CHO)-K1 cell mutants at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus were isolated from untreated, 60Co γ ray-and 212Bi α-exposed cells and the genetic changes underlying the mutation determined by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. In the 71 spontaneous mutants analyzed, 77.5% of the clones showed no change in exon number or size, 15.5% showed a loss of a single exon, 4.2% showed a loss of 2-8 exons, and 2.8% showed loss of all nine hprt exons (total gene deletion). Exposure to 6 Gy of γ rays, which reduced survival levels to 10%, produced a significantly different deletion spectrum that was shifted toward deletions with 45% of the 20 mutants analyzed showing a loss of a single exon and 30% showing a loss of all nine exons. Exposure to 2 Gy α radiation from 212Bi, a 220Rn daughter, a dose which also reduced survival levels to about 10%, resulted in a deletion spectrum similar to the γ-ray spectrum in that more than 75% of the 49 mutants analyzed were deletions. The α spectrum, however, was significantly different from both the spontaneous and γ spectra with 55.1% of the α mutants showing a loss of all nine exons, 10.2% showing loss of a single exon, and 14.3% showing loss of 2-8 exons. Thus, α-radiation appears to produce larger intragenic deletions than γ radiation. The results suggest that intragenic deletion size should be considered when low- and high linear energy transfer (LET) mutation spectra are compared.

AB - Independent Chinese hamster ovary (CHO)-K1 cell mutants at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus were isolated from untreated, 60Co γ ray-and 212Bi α-exposed cells and the genetic changes underlying the mutation determined by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. In the 71 spontaneous mutants analyzed, 77.5% of the clones showed no change in exon number or size, 15.5% showed a loss of a single exon, 4.2% showed a loss of 2-8 exons, and 2.8% showed loss of all nine hprt exons (total gene deletion). Exposure to 6 Gy of γ rays, which reduced survival levels to 10%, produced a significantly different deletion spectrum that was shifted toward deletions with 45% of the 20 mutants analyzed showing a loss of a single exon and 30% showing a loss of all nine exons. Exposure to 2 Gy α radiation from 212Bi, a 220Rn daughter, a dose which also reduced survival levels to about 10%, resulted in a deletion spectrum similar to the γ-ray spectrum in that more than 75% of the 49 mutants analyzed were deletions. The α spectrum, however, was significantly different from both the spontaneous and γ spectra with 55.1% of the α mutants showing a loss of all nine exons, 10.2% showing loss of a single exon, and 14.3% showing loss of 2-8 exons. Thus, α-radiation appears to produce larger intragenic deletions than γ radiation. The results suggest that intragenic deletion size should be considered when low- and high linear energy transfer (LET) mutation spectra are compared.

UR - http://www.scopus.com/inward/record.url?scp=0028113048&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028113048&partnerID=8YFLogxK

U2 - 10.1093/mutage/9.6.537

DO - 10.1093/mutage/9.6.537

M3 - Article

VL - 9

SP - 537

EP - 540

JO - Mutagenesis

JF - Mutagenesis

SN - 0267-8357

IS - 6

ER -