CRE RECOMBINASE-MEDIATED SITE-SPECIFIC RECOMBINATION BETWEEN PLANT CHROMOSOMES
CRE RECOMBINASE-MEDIATED SITE-SPECIFIC RECOMBINATION BETWEEN PLANT CHROMOSOMES
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DOI：
10.1073/pnas.91.5.1706
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发表时间：
1994-03-01
影响因子：
11.1
通讯作者：
OW, DW
中科院分区：
文献类型：
--
作者：
QIN, MM;BAYLEY, C;OW, DW
我们报道了使用噬菌体P1 Cre-lox系统在烟草染色体之间产生保守的位点特异性重组。将两种构建物分别导入烟草植株，其中一种构建物含有一个lex位点前的无启动子抗潮霉素基因（lox-hpt），另一种构建物含有一个连接fox序列和cre编码区的花菜花叶病毒35S启动子（35S-lox-cre）。含有任意一种结构的植物之间的杂交产生了位于不同染色体上的两种结构的植物。通过选择重组后表达的抗潮霉素表型来鉴定具有重组事件的植株。分子分析表明，这些重组事件特异性地发生在lox位点，并导致两侧宿主DNA的相互交换。新基因35S-lox-hpt和lox-cre的共传率为67-100%，与易位染色体的优先共分离一致。这些结果表明，位点特异性重组系统可以成为真核生物体内染色体大规模操作的有用工具。
We report the use of the bacteriophage P1 Cre-lox system for generating conservative site-specific recombination between tobacco chromosomes. Two constructs, one containing a promoterless hygromycin-resistance gene preceded by a lex site (lox-hpt) and the other containing a cauliflower mosaic virus 35S promoter linked to a fox sequence and the cre coding region (35S-lox-cre), were introduced separately into tobacco plants. Crosses between plants harboring either construct produced plants with the two constructs situated on different chromosomes. Plants with recombination events were identified by selecting for hygromycin resistance, a phenotype expressed upon recombination. Molecular analysis showed that these recombination events occurred specifically at the lox sites and resulted in the reciprocal exchange of flanking host DNA. Progenies of these plants showed 67-100% cotransmission of the new transgenes, 35S-lox-hpt and lox-cre, consistent with the preferential cosegregation of translocated chromosomes. These results illustrate that site-specific recombination systems can be useful tools for the large-scale manipulation of eukaryotic chromosomes in vivo.