Preliminary Screening of Rice Promoter Trap Lines with T-DNA Tagging
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摘要: 利用农杆菌介导法将含报道基因β-glucuronidase(GUS)无启动子的转化载体pCAMBIA1300GUSA-Hyg导入籼稻明恢86获得水稻启动子捕获系。水稻启动子捕获系潮酶素抗感分离比χ2分析表明:39个水稻启动子捕获系中有28个为单T-DNA位点插入。2653个水稻启动子捕获系报告基因GUS表达检测结果表明:83个水稻启动子捕获系在其不同时期、不同器官(包括根、茎、叶、花药、颖壳、胚、胚乳等组织)中检测到GUS表达。其中GUS活性在胚乳有表达的水稻启动子捕获系24个,占检测总数0.91%;GUS活性在花药中有表达的水稻启动子捕获系为19个,占检测总数0.72%。Abstract: Binary vector pCAMBIA1300GUSA-Hyg containing β-glucuronidase(GUS) without the promoter was introduced into Oryza sativa L.Minghui 86 by agrobacterium mediated transformation.The χ2 tests on the hygromycin resistant ratio revealed that 28 of the 39 rice promoter trap lines contained a loci T-DNA tagging.GUS histochemical assay at different stages in different organs,such as root,stem,leave,anther,hull,embryo and emdospern,of 2653 rice promoter trap lines was performed.The results showed that 83 rice promoter lines were GUS positive.Of all tested,24 lines were expressed in endosperm(i.e.,0.91%) and 19 lines expressed in anther(i.e.,0.72%).Information collected on the rice promoter trap lines could be useful in identifying new rice genes.
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Key words:
- Rice /
- promoter trap lines /
- T-DNA tagging
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[1] JEON JS,LEE S,JUNG KH,et al.T-DNA insertional mutagenesis for functional genomics in rice[J].Plant J,2000,22 (6):561-70. [2] SPRINGER PS.Gene traps:tool for plant development and genomics[J].Plant Cell,2000,12:1007-1020. [3] JEFFERSON R A.Assaying chimeric genes in plants:the GUS gene fusion system[J].Plant Molecular Biology Reporter,1987,5:387. [4] ZHANG J,LI C,WU C,et al.RMD:a rice mutant database for functional analysis of the rice genome[J].Nucleic Acids Res,2006,1 (34):745-748.
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