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采用MADS(MCM1-Agamous-Deficiens-SRF)盒基因家族功能区保守序列PCR引物,将水稻(Oryzasativa L.ssp indica)“珍汕97B”悬浮细胞、愈伤组织、分化愈伤组织和再生试管苗等不同形态发生的组织的mRNA反转录后选择性放大,经测序胶分离鉴定出一组差异表达的cDNA。对命名为RM1 cDNA的5＇端序列测定表明,RM1与典型的MADS基因——拟南芥agamous蛋白保守区一级结构同源性达63％,模拟二级结构相似性显著,初步确认RM1属于MADS基因家族成员。分子杂交证实,RM1在悬浮培养细胞中不表达,而在愈伤组织、分化愈伤组织和再生试管苗中活跃表达。
The rice (Oryzasativa L.ssp indica) “Zhenshan 97B” suspension cells, callus, differentiated calli and regeneration test-tube seedlings were prepared by using the conserved sequence PCR primers of MADS (MCM1-Agamous-Deficiens- Such as different morphological changes of mRNA mRNA synthesis after selective amplification, isolated by sequencing gel to identify a group of differentially expressed cDNA. The 5 ’end sequence of RM1 cDNA showed that RM1 had 63% primary structure homology with the typical MADS gene - Arabidopsis thaliana agamous protein. The similarity of the simulated secondary structure was significant. It was initially confirmed that RM1 belonged to MADS gene family members. Molecular hybridization confirmed that RM1 was not expressed in suspension cultured cells but was actively expressed in callus, differentiated callus and regenerated plantlets.