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以cyclAt基因为目的基因,HPT和GUS为选择和报告基因,应用PDS-1 000/He型基因枪协同转导,通过轰击来源于水稻成熟种子诱导产生的胚性愈伤组织,获得了转基因水稻植株.结果表明:通过X-Gluc染色分析,GUS基因在愈伤组织和再生植株叶片中的表达效率高.以转基因植株的基因组DNA为模板,用cyclAt基因的特异性引物对其进行PCR扩增,只检测到1 000 bp左右的片段,比其1 604 bp的完整序列小,因此推测cyc1At基因在转导入水稻后被剪接.此外,在协同转导过程中,cyc1At和GUS可能与受体基因组DNA随机整合,因此视GUS基因为报告基因有一定局限性;但两者同时被整合的频率远高于单一基因的整合.
The cloned cDNAs were cloned and sequenced. The cloned genes were cloned and sequenced. The cloned cDNAs were cloned and sequenced. The cloned cDNAs were cloned and sequenced. The genes of cyclAt gene, HPT and GUS were selected and reported. Synergistic transduction with PDS-1 000 / He gene gun was used to bombard the embryogenic callus induced from ripe seeds of rice. The results showed that the expression of GUS gene in leaves of callus and regenerated plant was highly efficient by X-Gluc staining.The genomic DNA of transgenic plants was used as a template to amplify the GUS gene with specific primers of cyclAt gene , Only about 1 000 bp fragment was detected, which is smaller than its complete sequence of 1 604 bp, suggesting that cyc1At gene is spliced after transfection into rice.In addition, cyc1At and GUS may interact with receptor genome Therefore, the GUS gene has some limitations as a reporter gene. However, the frequency of integration of the two genes is much higher than that of a single gene.