PUBLICATION DATE

2008-05-07

AUTHORS

Jianxin Niu, Lizhong Pan, Huping Zhang, Xiaoyu Duan, Binggang Ma, Chao Ma

TITLE

Salicylic-Acid-Induced Self-excision of the Marker Gene nptII from Transgenic Tomato Using the Cre–loxP System

ISSUE

3

VOLUME

26

ISSN (print)

0735-9640

ISSN (electronic)

1572-9818

ABSTRACT

The presence of antibiotic-resistant genes in genetically engineered crops together with the target gene has generated a number of environmental and consumer concerns. In order to alleviate public concerns over the safety of food derived from transgenic crops, marker gene elimination is desirable. Marker-free transgenic tomato plants were obtained by using a salicylic-acid-regulated Cre–loxP-mediated site-specific DNA recombination system in which the selectable marker neomycin phosphotransferase nptII and cre genes were flanked by two directly oriented loxP sites. Upon induction by salicylic acid, the cre gene produced a recombinase that eliminated sequences encoding nptII and cre genes, sandwiched by two loxP sites from the tomato genome. Regenerant plants with the Cre–loxP system were obtained by selection on kanamycin media and polymerase chain reaction (PCR) screening. Transgenic plants were screened for excision by PCR using nptII, cre, and PR-1a promoter primers following treatment with salicylic acid. The footprint of the excision was determined by sequencing the T-DNA borders after a perfect recombination event. The excision efficiency was 38.7%. A new plant transformation vector, pBLNSC (Genbank accession number EU327497), was developed, containing six cloning sites and the self-excision system. This provided an effective approach to eliminate the selectable marker gene from transgenic tomato, thus expediting public acceptance of genetically modified tomato.

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1 articles:4c150cb6024f17eb24b8295c70ab5e34 sg:abstract Abstract The presence of antibiotic-resistant genes in genetically engineered crops together with the target gene has generated a number of environmental and consumer concerns. In order to alleviate public concerns over the safety of food derived from transgenic crops, marker gene elimination is desirable. Marker-free transgenic tomato plants were obtained by using a salicylic-acid-regulated Cre–loxP-mediated site-specific DNA recombination system in which the selectable marker neomycin phosphotransferase nptII and cre genes were flanked by two directly oriented loxP sites. Upon induction by salicylic acid, the cre gene produced a recombinase that eliminated sequences encoding nptII and cre genes, sandwiched by two loxP sites from the tomato genome. Regenerant plants with the Cre–loxP system were obtained by selection on kanamycin media and polymerase chain reaction (PCR) screening. Transgenic plants were screened for excision by PCR using nptII, cre, and PR-1a promoter primers following treatment with salicylic acid. The footprint of the excision was determined by sequencing the T-DNA borders after a perfect recombination event. The excision efficiency was 38.7%. A new plant transformation vector, pBLNSC (Genbank accession number EU327497), was developed, containing six cloning sites and the self-excision system. This provided an effective approach to eliminate the selectable marker gene from transgenic tomato, thus expediting public acceptance of genetically modified tomato.
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7 sg:doi 10.1007/s11105-008-0039-2
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34 sg:scigraphId 4c150cb6024f17eb24b8295c70ab5e34
35 sg:title Salicylic-Acid-Induced Self-excision of the Marker Gene nptII from Transgenic Tomato Using the Cre–loxP System
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37 sg:webpage https://link.springer.com/10.1007/s11105-008-0039-2
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39 rdfs:label Article: Salicylic-Acid-Induced Self-excision of the Marker Gene nptII from Transgenic Tomato Using the Cre–loxP System
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