Insertion of a heterologous gene construct into a non-functional ORF of the Streptococcus thermophilus chromosome |
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Authors: | John A Renye and George A Somkuti |
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Institution: | (1) Department of Agriculture, Eastern Regional Research Center, Agricultural Research Service, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA |
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Abstract: | An integrative vector was constructed for inserting heterologous genes within a non-functional open reading frame (ORF) on
the chromosome of Streptococcus thermophilus. The vector, pINTRS, contained a temperature sensitive origin of replication and an erythromycin resistance gene for initial
selection in S. thermophilus. The region of the vector containing unique cloning sites, for insertion of recombinant genes, was flanked by homologous
DNA sequences corresponding to a pseudogene in S. thermophilus to facilitate chromosomal integration. The gene encoding green fluorescent protein, regulated by a plasmid borne hsp promoter of S. thermophilus, was cloned into pINTRS to demonstrate proper functioning of the vector.
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Keywords: | Streptococcus thermophilus Integrative plasmid Green fluorescent protein |
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