DIRECT PCR SEQUENCING OF THE NDD GENE OF BACTERIOPHAGE-T4 - IDENTIFICATION OF A PRODUCT INVOLVED IN BACTERIAL NUCLEOID DISRUPTION

被引:17
作者
BOUET, JY [1 ]
WOSZCZYK, J [1 ]
REPOILA, F [1 ]
FRANCOIS, V [1 ]
LOUARN, JM [1 ]
KRISCH, HM [1 ]
机构
[1] LAB MICROBIOL & GENET MOLEC,CNRS,UPR 9007,F-31062 TOULOUSE,FRANCE
关键词
SEQUENCING OF PCR FRAGMENTS; INSERTIONAL MUTAGENESIS; ANALYSIS OF TOXIC GENES;
D O I
10.1016/0378-1119(94)90121-X
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The rapid disruption of the Escherichia coli nucleoid after T4 infection requires the activity of the phage-encoded ndd gene. We have genetically identified the sequence encoding ndd. Determination of the sequence of a 2.5-kb segment including ndd closed the last significant gap in the sequence of the T4 genome. This analysis was performed on PCR-amplified fragments that were purified by gel-exclusion chromatography and then submitted to linear amplication cycle sequencing. This technology permitted sequence comparison of two ndd mutants (ndd44 and ndd98) with the wild-type gene. The analysis of ndd from six bacteriophages of the T-even family indicated that the protein encoded by this nonessential gene is surprisingly conserved.
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页码:9 / 16
页数:8
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