Modification of human β-globin locus PAC clones by homologous recombination in Escherichia coli

被引：46

作者：

Imam, A. M. Ali ^{[1
]}

Patrinos, George P. ^{[1
]}

de Krom, Mariken ^{[1
]}

Bottardi, Stefania ^{[1
]}

Janssens, Rick J. ^{[1
]}

Katsantoni, Eleni ^{[2
,3
]}

Wai, Albert W. K. ^{[1
]}

Sherratt, David J. ^{[4
]}

Grosveld, Frank G. ^{[1
]}

机构：

[1] Erasmus Univ, CBG & MGC Dept Cell Biol, NL-3000 DR Rotterdam, Netherlands

[2] Univ Crete, Fac Med, Iraklion 71110, Crete, Greece

[3] Inst Mol Biol & Biotechnol, Iraklion 71110, Crete, Greece

[4] Univ Oxford, Dept Biochem, Div Mol Genet, Oxford OX1 3QU, England

来源：

NUCLEIC ACIDS RESEARCH | 2000年 / 28卷 / 12期

关键词：

D O I：

10.1093/nar/28.12.e65

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：

We report here modifications of human beta-globin PAC clones by homologous recombination in Escherichia coli DH10B, utilising a plasmid temperature sensitive for replication, the recA gene and a wild-type copy of the rpsL gene which allows for an efficient selection for plasmid loss in this host. High frequencies of recombination are observed even with very small lengths of homology and the method has general utility for introducing insertions, deletions and point mutations. No rearrangements were detected with the exception of one highly repetitive genomic sequence when either the E. coli RecA- or the lambdoid phage encoded RecT and RecE-dependent recombination systems were used.

引用

页数：6

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