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Genome-wide cloning and sequence analysis of leucine-rich repeat receptor-like protein kinase genes in Arabidopsis thaliana

机译:拟南芥中富含亮氨酸的重复受体样蛋白激酶基因的全基因组克隆和序列分析

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Background Transmembrane receptor kinases play critical roles in both animal and plant signaling pathways regulating growth, development, differentiation, cell death, and pathogenic defense responses. In Arabidopsis thaliana, there are at least 223 Leucine-rich repeat receptor-like kinases (LRR-RLKs), representing one of the largest protein families. Although functional roles for a handful of LRR-RLKs have been revealed, the functions of the majority of members in this protein family have not been elucidated. Results As a resource for the in-depth analysis of this important protein family, the complementary DNA sequences (cDNAs) of 194 LRR-RLKs were cloned into the GatewayR donor vector pDONR/ZeoR and analyzed by DNA sequencing. Among them, 157 clones showed sequences identical to the predictions in the Arabidopsis sequence resource, TAIR8. The other 37 cDNAs showed gene structures distinct from the predictions of TAIR8, which was mainly caused by alternative splicing of pre-mRNA. Most of the genes have been further cloned into GatewayR destination vectors with GFP or FLAG epitope tags and have been transformed into Arabidopsis for in planta functional analysis. All clones from this study have been submitted to the Arabidopsis Biological Resource Center (ABRC) at Ohio State University for full accessibility by the Arabidopsis research community. Conclusions Most of the Arabidopsis LRR-RLK genes have been isolated and the sequence analysis showed a number of alternatively spliced variants. The generated resources, including cDNA entry clones, expression constructs and transgenic plants, will facilitate further functional analysis of the members of this important gene family.
机译:背景跨膜受体激酶在调节生长,发育,分化,细胞死亡和致病性防御反应的动植物信号途径中都起着关键作用。在拟南芥中,至少有223种富含亮氨酸的重复受体样激酶(LRR-RLK),代表最大的蛋白质家族之一。尽管已经揭示了少数LRR-RLK的功能角色,但尚未阐明该蛋白家族中大多数成员的功能。结果作为深入分析此重要蛋白质家族的资源,将194个LRR-RLK的互补DNA序列(cDNA)克隆到Gateway R 供体载体pDONR / Zeo R中并通过DNA测序进行分析。其中,有157个克隆显示的序列与拟南芥序列资源TAIR8中的预测相同。其他37个cDNA显示的基因结构不同于TAIR8的预测,这主要是由pre-mRNA的可变剪接引起的。大多数基因已进一步克隆到具有GFP或FLAG表位标签的Gateway R 目的地载体中,并已转化到拟南芥中以进行植物功能分析。这项研究的所有克隆均已提交给俄亥俄州立大学的拟南芥生物资源中心(ABRC),以供拟南芥研究界全面访问。结论大部分拟南芥LRR-RLK基因已被分离,序列分析显示了许多剪接的变异体。产生的资源,包括cDNA进入克隆,表达构建体和转基因植物,将有助于对该重要基因家族成员的进一步功能分析。

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