New mutation detection system of repackaged λ gt11 DNA containing LacZ gene
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New mutation detection system of repackaged λ gt11 DNA containing LacZ gene
Military Medical ResearchIssue 3, Pages: 162-166(2002)
Affiliations:
1. Department of Hygiene Toxicology
2. Third Military Medical University
3. ,China
Author bio:
Funds:
DOI:
CLC:R346
Published:2002
Accepted:
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[1]刘勇,曹佳,吴涛,杨录军,孙华明,杨明杰,钱频.New mutation detection system of repackaged λ gt11 DNA containing LacZ gene[J].Journal of Medical Colleges of PLA,2002(03):162-166.
DOI:
[1]刘勇,曹佳,吴涛,杨录军,孙华明,杨明杰,钱频.New mutation detection system of repackaged λ gt11 DNA containing LacZ gene[J].Journal of Medical Colleges of PLA,2002(03):162-166.DOI:
New mutation detection system of repackaged λ gt11 DNA containing LacZ gene
摘要
Abstract
<正> Objective: To establish a reformative detection system which has sound ability of providing information on molecular mutagenesis spectrum and the specificity of detection system of repackaged λ phage. Methods: LacZ gene
as mutational target gene and reporter gene
was applied into the detection system. The λ gt11 DNA treated with ENU (1-ethyl-1-nitrosourea) and 9-AA (9-aminoacridine) was repackaged in vitro. The packaged λ phage was then grown in E. coli Y1090 on a selective plate containing X-gel and IPTG. The survival and mutation frequencies were determined by counting the clear-plaque and blue-plaque
and the molecular mutation mechanism was studied by extracting and sequencing the LacZ gene of mutants. Results: The survival of repackaged λ phages treated with 9-AA and ENU apparently decreased in consistent dose-dependence. The mutation frequency of clear-plaque mutants showed a linear dose-related increase. The predominant mutations induced by 9-AA were ± 1 frameshift mutation
and 9-AA induc
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