Cloning of antibiotic-resistance genes from Streptomyces and analysis of their expression.
| Project/Area Number |
59480412
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Biological pharmacy
|
|---|
| Research Institution | Meiji College of Pharmacy |
|---|
Principal Investigator |
OGAWARA Hiroshi Meiji College of Pharmacy, 薬学部, 教授 (00097198)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NAKANO Michiko Meiji College of Pharmacy, 薬学部, 講師 (60125086)
|
|---|
| Project Period (FY) |
1984 – 1986
|
| Project Status |
Completed (Fiscal Year 1986)
|
| Budget Amount *help |
¥6,200,000 (Direct Cost: ¥6,200,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1985: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1984: ¥3,800,000 (Direct Cost: ¥3,800,000)
|
|---|
| Keywords | Streptomyces / Cloning of antibiotic resistance genes / Gene expression / プロモーター |
|---|
| Research Abstract |
1. Kanamycin resistance gene
A Kanamycin resistance gene (kan) was cloned from Streptomyces kanamyceticus. It was shown that 30S subunit confers resistance on ribosomes of the transformants. While the expression of the kan gene in S. kanamyceticus was inducible under the same regulation with that of the kanamycin production, it was constitutively expressed in the cells carrying the cloned kan gene on plasmid (both multi-copy and low-copy ones).
High-resolution Sl mapping identified the putative transcription start site of the kan gene on plasmid. The kan promoter showed significant homology with those of vphPl and endoH in -10 region. Another transcript, using the opposite strand as the template, was detected and the transcription start site was identified to be several base pairs downstream from the start point of kan mRNA.
The transcription of the kan gene was examined in the donor strain, S. kanamyceticus. The kan mRNA was not transcribed under the condition which repressed kanamycin production, showing that the regulation of the gene expression is transcriptional level. Under the condition which allowed kanamycin production, the kan gene was transcribed, however, the strat point was different from that of the gene cloned on plasmid DNA.
2. Penicillinase gene
penicillinase gene cloned from S. cacaoi was identified as 2 kb fragment. DNA sequencing of the fragment suggested probable open reading frame, showing homologous amino acid sequence with penicillinase genes from other bacteria.
Using promoter-probe vector and Sl mapping, promoter region of the gene was identified.
|
Report
(1 results)
Research Products
(12 results)
