Abstract
A Tn3 derivative was constructed to make small in-frame insertions within genes. The transposon contains the URA3 gene, the tetA gene, a truncated lacZ, and phage P1 loxP recombination sites at either end. Insertions that have fused lacZ to an open reading frame are lac+ because they express the truncated lacZ. In the presence of the phage P1 cyclization recombinase cre, the transposon can delete the URA3, tetA, and lacZ genes between the two loxP sites. The remaining short imperfect palindrome contains the ends of Tn3 and a loxP site and does not contain a translational termination codon in the correct reading frame. We have analyzed several insertions within the yeast HO gene. Several insertions inactivate HO and prohibit initiation of mating-type switching. In contrast, an epitope inserted in the central portion encodes a functional HO endonuclease.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 5457-5461 |
| Number of pages | 5 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 88 |
| Issue number | 12 |
| State | Published - Jun 15 1991 |
| Externally published | Yes |
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Keywords
- Saccharomyces cerevisiae
- Transposon mutagenesis
ASJC Scopus subject areas
- General
- Genetics
Cite this
A Tn3 derivative that can be used to make short in-frame insertions within genes. / Hoekstra, M. F.; Burbee, D.; Singer, J.; Mull, E.; Chiao, E.; Heffron, Fred.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 88, No. 12, 15.06.1991, p. 5457-5461.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A Tn3 derivative that can be used to make short in-frame insertions within genes
AU - Hoekstra, M. F.
AU - Burbee, D.
AU - Singer, J.
AU - Mull, E.
AU - Chiao, E.
AU - Heffron, Fred
PY - 1991/6/15
Y1 - 1991/6/15
N2 - A Tn3 derivative was constructed to make small in-frame insertions within genes. The transposon contains the URA3 gene, the tetA gene, a truncated lacZ, and phage P1 loxP recombination sites at either end. Insertions that have fused lacZ to an open reading frame are lac+ because they express the truncated lacZ. In the presence of the phage P1 cyclization recombinase cre, the transposon can delete the URA3, tetA, and lacZ genes between the two loxP sites. The remaining short imperfect palindrome contains the ends of Tn3 and a loxP site and does not contain a translational termination codon in the correct reading frame. We have analyzed several insertions within the yeast HO gene. Several insertions inactivate HO and prohibit initiation of mating-type switching. In contrast, an epitope inserted in the central portion encodes a functional HO endonuclease.
AB - A Tn3 derivative was constructed to make small in-frame insertions within genes. The transposon contains the URA3 gene, the tetA gene, a truncated lacZ, and phage P1 loxP recombination sites at either end. Insertions that have fused lacZ to an open reading frame are lac+ because they express the truncated lacZ. In the presence of the phage P1 cyclization recombinase cre, the transposon can delete the URA3, tetA, and lacZ genes between the two loxP sites. The remaining short imperfect palindrome contains the ends of Tn3 and a loxP site and does not contain a translational termination codon in the correct reading frame. We have analyzed several insertions within the yeast HO gene. Several insertions inactivate HO and prohibit initiation of mating-type switching. In contrast, an epitope inserted in the central portion encodes a functional HO endonuclease.
KW - Saccharomyces cerevisiae
KW - Transposon mutagenesis
UR - http://www.scopus.com/inward/record.url?scp=0026077655&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026077655&partnerID=8YFLogxK
M3 - Article
C2 - 1647034
AN - SCOPUS:0026077655
VL - 88
SP - 5457
EP - 5461
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 12
ER -