Inviable phages, their production and DNA thereof

Details Inviable phages, their production and DNA thereof Inviable phages, their production and DNA thereof

1995-06-06

1998-11-10

Martinell, James

Chemistry: molecular biology and microbiology

Virus or bacteriophage, except for viral vector or...

4353201, 435 6, 536 231, C12N 1500, C12N 1511, C12Q 168

Patent

active

058342915

ABSTRACT:
Inviable T4 phage-like particles capable of directing the expression of large non-T4 DNA fragments from T4 expression control sequences are produced. Thus, E. coli harboring pBR322 derivatives containing cloned T4 gene 23 DNA sequences were infected with T4 phage carrying a deletion of the denB gene. Homology-dependent recombination results in the production of inviable phage-like particles containing DNA molecules composed of multiple, tandemly repeated copies of entire plasmid molecules covalently linked to single copies of normal phage genes. The yield of these inviable particles, intially low, was increased by means of a reiterated infection process that involves the use of a cloned T4 origin of replication. When T4 gene 32 expression control sequences linked in proper orientation to a DNA sequence coding for the non-T4 protein .beta.-galactosidase were also cloned in one such pBR322 derivative (pVH773), inviable phage particles capable of directing the synthesis of enzymatically active .beta.-galactosidase were produced. The present process is applicable to other T-even bacterio-phages.

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