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Author:
Date:
2015-09-20
[Abstract] The T-DNA (transferred-DNA) region of virulent Agrobacterium tumefaciens (A. tumefaciens) strain is transferred and integrated into the plant genome, and thereby the T-DNA genes are expressed in transformed plant cells. This protocol was used to analyze the transcription start sites (TSSs) of agrobacterial T-DNA genes within plant crown gall tumor. Firstly, the stems of Arabidopsis thaliana were inoculated by A. tumefaciens strain C58 and developed crown gall tumor. Subsequently, the mRNA was extracted from the crown gall tumor and then used for amplification of 5’ cDNA ends by 5’ Rapid Amplification of cDNA Ends (5’ RACE) assay. The full-length cDNAs were generated in reverse transcription reactions and used to analyze TSSs. Here, TSSs of three ...
[摘要] 将有毒的根癌土壤杆菌（根瘤土壤杆菌）菌株的T-DNA（转移的DNA）区转移并整合到植物基因组中，从而T-DNA基因在转化的植物细胞中表达。该方案用于分析植物冠gall肿瘤内农杆菌T-DNA基因的转录起始位点（TSS）。首先，通过接种拟南芥的茎。 tumefaciens 菌株C58和发展的冠gall肿瘤。随后，从冠gall瘤提取mRNA，然后通过cDNA末端的快速扩增（5'RACE）测定用于扩增5'cDNA末端。全长cDNA在逆转录反应中产生并用于分析TSS。这里，作为实例分析了三种癌基因的TSS，IaaH ，IaaM 和 Ipt 。该协议还允许鉴定在植物细胞中表达的其他农杆菌T-DNA基因的TSS
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