Transient expression of a foreign gene by direct incorporation of DNA into intact plant tissue through vacuum infiltration
Transient expression of a foreign gene by direct incorporation of DNA into intact plant tissue through vacuum infiltration
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DOI：
10.1007/s10529-009-0080-8
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发表时间：
2009-07
影响因子：
2.7
通讯作者：
Bo Zhou;Xia Zhao;S. Kawabata;Yuhua Li
中科院分区：
文献类型：
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作者：
Bo Zhou;Xia Zhao;S. Kawabata;Yuhua Li
我们以前建立了一种方法，诱导外源基因在完整的植物组织中瞬时表达，以检测蛋白质的亚细胞定位。在这里，我们已经插入了一个推定的bZIP蛋白HY 5基因（SeqID：EU 386772），分离自芜菁幼苗。亚种从拟南芥中分离到的类受体激酶基因AtRLK（SeqID：AY531551.1）克隆到质粒pA 7-GFP中。采用真空渗入法将DNA直接导入洋葱表皮组织中。通过检测与AtRLK或推定的BrHY 5融合的GFP，我们确定BrHY 5位于细胞核中，AtRLK位于质膜中。因此，该方法可用于研究外源基因在完整组织中的瞬时表达。
We previously established a method to induce transient expression of foreign genes in intact plant tissue to detect the subcellular localization of proteins. Here, we have inserted a putative bZIP protein HY5 gene (SeqID: EU386772), isolated from the seedlings of turnipsBrassica rapaL. subsp. rapa ‘Tsuda,’ and a receptor-like kinase gene AtRLK (SeqID: AY531551.1), isolated fromArabidopsis, into the plasmid pA7-GFP. We accomplished the direct incorporation of DNA into onion epidermal tissue by vacuum infiltration. By detecting GFP, which was fused with AtRLK or putative BrHY5, we determined that BrHY5 is located in the nucleus and AtRLK is located in the plasma membrane. This approach can be thus used to study the transient expression of foreign genes in intact tissue.