Cre-lox univector acceptor vectors for functional screening in protoplasts: analysis of Arabidopsis donor cDNAs encoding ABSCISIC ACID INSENSITIVE1-like protein phosphatases.
Cre-lox univector acceptor vectors for functional screening in protoplasts: analysis of Arabidopsis donor cDNAs encoding ABSCISIC ACID INSENSITIVE1-like protein phosphatases.
复制标题
用于原生质体功能筛选的 Cre-lox univector 受体载体：编码 ABSCISIC ACID INSENSITIVE1 样蛋白磷酸酶的拟南芥供体 cDNA 分析。
DOI：
10.1007/s11103-009-9502-1
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发表时间：
2009
影响因子：
5.1
通讯作者：
Rock,ChristopherD
中科院分区：
文献类型：
--
作者：
Jia,Fan;Gampala,SrinivasSL;Mittal,Amandeep;Luo,Qingjun;Rock,ChristopherD
通用质粒系统(UPS)供体载体pUNI51中14,200个可利用的全长拟南芥DNA应该被广泛和有效地应用于利用同源植物基因的“功能图谱空间”。我们已经构建了Cre-lox UPS宿主受体载体(pCR701-705)，其N端表位标签位于loxH位点的框架内，位于玉米泛素启动子的下游，用于原生质体瞬时表达分析和单子叶植物的基因轰击转化。作为这些载体用途的一个例子，我们将它们与几个编码丝氨酸/苏氨酸蛋白磷酸酶2C(PP2Cs)的拟南芥DNA重组，这些DNA来自遗传学研究或通过等级聚类荟萃分析预测与ABA和胁迫反应有关。我们在玉米可能叶原生质体中对ABA诱导表达对胚胎发育后期丰富的启动子PROEM：GUS的功能结果与预测一致，并鉴定了一些PP2Cs的新活性。这些载体的部署可以促进功能基因组学和蛋白质组学以及新基因活性的鉴定。
The 14,200 available full lengthArabidopsis thalianacDNAs in the universal plasmid system (UPS) donor vector pUNI51 should be applied broadly and efficiently to leverage a “functional map-space” of homologous plant genes. We have engineered Cre-lox UPS host acceptor vectors (pCR701- 705) with N-terminal epitope tags in frame with the loxH site and downstream from the maizeUbiquitinpromoter for use in transient protoplast expression assays and particle bombardment transformation of monocots. As an example of the utility of these vectors, we recombined them with severalArabidopsiscDNAs encoding Ser/Thr protein phosphatase type 2C (PP2Cs) known from genetic studies or predicted by hierarchical clustering meta-analysis to be involved in ABA and stress responses. Our functional results inZea maysmesophyll protoplasts on ABA-inducible expression effects on the Late Embryogenesis Abundant promoter ProEm:GUS reporter were consistent with predictions and resulted in identification of novel activities of some PP2Cs. Deployment of these vectors can facilitate functional genomics and proteomics and identification of novel gene activities.