Marker-free Transgenic Plants Produced by Autoexcised Cre-lox System
2007
Lee, W.S. (Kyung Hee University, Yongin, Republic of Korea) | Kim, S.H. (Kyung Hee University, Yongin, Republic of Korea) | Park, Y.D. (Kyung Hee University, Yongin, Republic of Korea), E-mail: [email protected]
Autoexcision strategy was investigated to remove an nptⅡ marker gene from transgenic tobacco. In the autoexcision strategy, the cre gene, under the control of a heat shock-inducible promoter, was excised along with the nptⅡ marker gene. Although no direct relation can be proven between the phenotype and cre expression in some plants, expressing the cre gene driven by a CaMV 35S promoter displayed growth retardation and a distinct pattern of chlorosis in the leaves. The recombinant event could be detected because it places the CaMV 35S promoter of the nptⅡ gene adjacent to a promoterless gusA gene. As a result the gusA gene was activated and its expression could be visualized. In the autoexcision strategy, 20 lines showed low or even could not detect GUS activity before heat treatment. However, after heat treatment, the degree of GUS expression increased in 16 lines after heat treatment. In this case, recombination occurred as high efficiency. Our results show that a transient heat shock treatment is sufficient for inducing cre and excising the cre and nptⅡ gene. Molecular analysis and GUS assay confirmed that marker gene removal was precise, complete and stable.
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