| 摘要 |
Hexulose phosphate synthase was purified from Mycobacterium gastri to determine an amino acid sequence of the enzyme. Oligonucleotide primers to be used for PCR were synthesized on the basis of obtained amino acid sequence information. A DNA fragment, which was amplified by performing PCR by using a template of genomic DNA prepared from Mycobacterium gastri, was used as a probe so that colony hybridization was carried out with respect to a library comprising fragments obtained by digesting Mycobacterium gastri genomic DNA with PstI to obtain a positive clone. Thus, the DNA fragment which contains ORF encoding hexulose phosphate synthase was obtained. Furthermore, the DNA fragment cloned contained other ORF(ORF-1). The expression system of ORF-1 was constructed (pT-HPIS-1), and it was introduced into Escherichia coli cell. After the induction of the expression of ORF-1, the hexulose phosphate isomerase activity was found in the extract of cells harvoring pT-HPIS-1. Thus, gene, which coded for hexulose phosphate isomerase, was obtained together with ORF which coded for hexulose phosphate synthase.
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