Abstract
The hybrid plasmid pYT72/pYT92 constructed from an Escherichia coli plasmid pACYC177 and mycobacterial plasmid pMSC262 isolated from Mycobacterium scroflaceum strain W262 transformed both E. coli and BCG. Phage-sensitive mutants S-10 and S-20 isolated from BCG Tokyo strain showed higher frequency of transformation than the wild-type strain. Frequency of transformation was dependent on age of the culture and the electroporation condition used. Several deletion mutants were generated from pYT72/92 to determine the minimum region for the replication in the mycobacteria. A 2.3-kb fragment of pMSC262 was found to contain an essential region. Using this fragment and pACYC177, a small shuttle vector pYT937 containing two drug-resistance markers, kanamycin- and ampicillin-resistance, was constructed. pYT937 contains AatII, BamHI, BbvII, GsuI, HincII, PstI, ScaI and XbaI cloning sites.
| Original language | English |
|---|---|
| Pages (from-to) | 277-282 |
| Number of pages | 6 |
| Journal | FEMS microbiology letters |
| Volume | 83 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - Oct 15 1991 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Microbiology
- Molecular Biology
- Genetics
Cite this
Development of a new host vector system in mycobacteria. / Goto, Yoshitaka; Taniguchi, Hatsumi; Udou, Takezo; Mizuguchi, Yasuo; Tokunaga, Tohru.
In: FEMS microbiology letters, Vol. 83, No. 3, 15.10.1991, p. 277-282.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Development of a new host vector system in mycobacteria
AU - Goto, Yoshitaka
AU - Taniguchi, Hatsumi
AU - Udou, Takezo
AU - Mizuguchi, Yasuo
AU - Tokunaga, Tohru
PY - 1991/10/15
Y1 - 1991/10/15
N2 - The hybrid plasmid pYT72/pYT92 constructed from an Escherichia coli plasmid pACYC177 and mycobacterial plasmid pMSC262 isolated from Mycobacterium scroflaceum strain W262 transformed both E. coli and BCG. Phage-sensitive mutants S-10 and S-20 isolated from BCG Tokyo strain showed higher frequency of transformation than the wild-type strain. Frequency of transformation was dependent on age of the culture and the electroporation condition used. Several deletion mutants were generated from pYT72/92 to determine the minimum region for the replication in the mycobacteria. A 2.3-kb fragment of pMSC262 was found to contain an essential region. Using this fragment and pACYC177, a small shuttle vector pYT937 containing two drug-resistance markers, kanamycin- and ampicillin-resistance, was constructed. pYT937 contains AatII, BamHI, BbvII, GsuI, HincII, PstI, ScaI and XbaI cloning sites.
AB - The hybrid plasmid pYT72/pYT92 constructed from an Escherichia coli plasmid pACYC177 and mycobacterial plasmid pMSC262 isolated from Mycobacterium scroflaceum strain W262 transformed both E. coli and BCG. Phage-sensitive mutants S-10 and S-20 isolated from BCG Tokyo strain showed higher frequency of transformation than the wild-type strain. Frequency of transformation was dependent on age of the culture and the electroporation condition used. Several deletion mutants were generated from pYT72/92 to determine the minimum region for the replication in the mycobacteria. A 2.3-kb fragment of pMSC262 was found to contain an essential region. Using this fragment and pACYC177, a small shuttle vector pYT937 containing two drug-resistance markers, kanamycin- and ampicillin-resistance, was constructed. pYT937 contains AatII, BamHI, BbvII, GsuI, HincII, PstI, ScaI and XbaI cloning sites.
UR - http://www.scopus.com/inward/record.url?scp=0026425713&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026425713&partnerID=8YFLogxK
U2 - 10.1111/j.1574-6968.1991.tb04477.x
DO - 10.1111/j.1574-6968.1991.tb04477.x
M3 - Article
C2 - 1769534
AN - SCOPUS:0026425713
VL - 83
SP - 277
EP - 282
JO - FEMS Microbiology Letters
JF - FEMS Microbiology Letters
SN - 0378-1097
IS - 3
ER -