发明名称 |
Method for constructing a transposon for identifying a DNA sequence coding for a signal peptide in lactic acid bacteria |
摘要 |
A method for constructing a transposon derivative for identifying in a lactic acid bacterium a DNA sequence coding for a signal peptide (SP) comprises selecting a DNA molecule comprising a transposon including between its left terminus (LR) and its right terminus (RR) a sequence comprising a promoterless promoter reporter gene and a ribosome binding site (RBS). A region located between the LR and the promoterless reporter gene is deleted form the DNA molecule to obtain a modified DNA molecule that has retained its transposability and its RBS. A unique restriction is then inserted into the remaining region located between the LR and the promoterless gene of the modified DNA molecule. A DNA sequence coding for a secretion reporter molecule, which is without a sequence coding for a SP, is inserted into the unique restriction site. The obtained transposon derivative is without stop codons in-frame with the reporter molecule thus allowing upon transposition translational fusions from upstream the LR. Also described is a method for identifying in a lactic acid bacteria a DNA coding sequence coding for a SP using the transposon derivative and the isolation of the sequences.
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申请公布号 |
NZ517079(A) |
申请公布日期 |
2003.08.29 |
申请号 |
NZ20000517079 |
申请日期 |
2000.08.04 |
申请人 |
BIOTEKNOLOGISK INSTITUT |
发明人 |
MADSEN, MICHAEL SOEREN;RAVN, PETER;VRANG, ASTRID;ISRAELSEN, HANS;JOHNSEN, MADS GROENVOLD;BREDMOSE, LARS;ARNAU, JOSE |
分类号 |
C12N15/09;C12N1/21;C12N15/10;C12N15/62;C12N15/74;C12Q1/34;C12R1/01;C12R1/225;(IPC1-7):C12N15/74 |
主分类号 |
C12N15/09 |
代理机构 |
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代理人 |
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主权项 |
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地址 |
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