FACILITATED ISOLATION OF RARE RECOMBINANTS BY LIGASE CHAIN-REACTION - SELECTION FOR INTRAGENIC CROSSOVER EVENTS IN THE DROSOPHILA OPTOMOTOR BLIND GENE

被引:1
作者
BALLES, J [1 ]
PFLUGFELDER, GO [1 ]
机构
[1] UNIV WURZBURG, LEHRSTUHL GENET, BIOZENTRUM, THEODOR BOVERI INST, D-97074 WURZBURG, GERMANY
来源
MOLECULAR AND GENERAL GENETICS | 1994年 / 245卷 / 06期
关键词
LIGASE CHAIN REACTION; INTRAGENIC RECOMBINATION; GENE REGULATION; POINT MUTATION; DROSOPHILA MELANOGASTER;
D O I
10.1007/BF00297280
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Ligase chain reaction (LCR) was evaluated as a tool for the detection of point mutations. For the mutation studied, the specificity of the method is sufficient to detect the mutant allele in the presence of a 200-fold molar excess of the wild-type sequence. LCR was therefore employed in a genetic recombination experiment as a probe for a recessive lethal point mutation. LCR greatly facilitated the isolation of a rare recombinant originating from a crossover event in the 40 kb interval separating the lethal mutation and an enhancer trap insertion in the optomotor-blind locus. The recombinant will allow the study of gene control in situ, in a largely unperturbed regulatory environment.
引用
收藏
页码:734 / 740
页数:7
相关论文
共 32 条
  • [1] TARGETED GENE-MUTATIONS IN DROSOPHILA
    BALLINGER, DG
    BENZER, S
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (23) : 9402 - 9406
  • [3] Barany F, 1991, PCR Methods Appl, V1, P5, DOI 10.1101/gr.1.1.5
  • [4] P-ELEMENT-MEDIATED ENHANCER DETECTION - A VERSATILE METHOD TO STUDY DEVELOPMENT IN DROSOPHILA
    BELLEN, HJ
    OKANE, CJ
    WILSON, C
    GROSSNIKLAUS, U
    PEARSON, RK
    GEHRING, WJ
    [J]. GENES & DEVELOPMENT, 1989, 3 (09) : 1288 - 1300
  • [5] SEARCHING FOR PATTERN AND MUTATION IN THE DROSOPHILA GENOME WITH A P-LACZ VECTOR
    BIER, E
    VAESSIN, H
    SHEPHERD, S
    LEE, K
    MCCALL, K
    BARBEL, S
    ACKERMAN, L
    CARRETTO, R
    UEMURA, T
    GRELL, E
    JAN, LY
    JAN, YN
    [J]. GENES & DEVELOPMENT, 1989, 3 (09) : 1273 - 1287
  • [6] CHARACTERIZATION OF A DROSOPHILA PROTEIN ASSOCIATED WITH BOUNDARIES OF TRANSCRIPTIONALLY ACTIVE CHROMATIN
    CHAMPLIN, DT
    FRASCH, M
    SAUMWEBER, H
    LIS, JT
    [J]. GENES & DEVELOPMENT, 1991, 5 (09) : 1611 - 1621
  • [7] PREVENTION OF PRE-PCR MIS-PRIMING AND PRIMER DIMERIZATION IMPROVES LOW-COPY-NUMBER AMPLIFICATIONS
    CHOU, Q
    RUSSELL, M
    BIRCH, DE
    RAYMOND, J
    BLOCH, W
    [J]. NUCLEIC ACIDS RESEARCH, 1992, 20 (07) : 1717 - 1723
  • [8] A 5' ELEMENT OF THE CHICKEN BETA-GLOBIN DOMAIN SERVES AS AN INSULATOR IN HUMAN ERYTHROID-CELLS AND PROTECTS AGAINST POSITION EFFECT IN DROSOPHILA
    CHUNG, JH
    WHITELEY, M
    FELSENFELD, G
    [J]. CELL, 1993, 74 (03) : 505 - 514
  • [9] FREEMAN M, 1992, DEVELOPMENT, V116, P335
  • [10] A 43 KILOBASE COSMID-P TRANSPOSON RESCUES THE FS(1K10 MORPHOGENETIC LOCUS AND 3 ADJACENT DROSOPHILA DEVELOPMENTAL MUTANTS
    HAENLIN, M
    STELLER, H
    PIRROTTA, V
    MOHIER, E
    [J]. CELL, 1985, 40 (04) : 827 - 837