Profilin has recently been identified as an actin-binding protein in higher plants. A cDNA clone (designated Repro ) encoding profilin gene was isolated from rapeseed ( Brassica napus L. cv. canadian Tween) using RT-PCR technique. Sequence analysis showed 82% similarity to Zea mays L. ZmPro3 , 85% to Arabidopsis Ath PRF1, 82% to Nicotiana tabacum L. NTPRO, 81% to Oryza sativa L. profilin A. A new full-length cDNA was obtained by 5′-RACE and 3′-RACE techniques. Sequence analysis showed that the size of full-length cDNA is 672 bp which contains a major open reading frame of 134 amino acids, 5′ and 3′ untranslated regions and a long Poly (A) tail. Northern blot analysis showed that the profilin gene is a pollen and anther specific gene.
油菜profilin 基因的克隆和表达分析
叶秋 李旭锋 徐莺 王劲林 娟陈放
（四川大学生命科学学院，成都610064）
摘要：profilin是高等植物中的一种与肌动蛋白结合的蛋白。采用RT-PCR技术克隆了油菜 (Bra ssica napus L. cv. canadian tween)花粉中的一个369 bp的cDNA片段。序列分析结果表明,该cDNA与已报道的其他植物的profilin基因具有较高核酸序列同源性。与玉米( Zea mays L.) 基因同源性为82％, 拟南芥(Arabidopsis)基因同源性为85％,水稻( Oryza sativa L.)基因同源性为81％,烟草(Nicotiana tabacum L.)基因同源性为8 2％。结合5′RACE和3′RACE 技术,获得了全长cDNA,其为672 bp。该cDNA包含一个开放密码框,5′未翻译区和一个带有Poly (A) 的3′区域。Northern 杂交结果显示它主要在花粉和花药中表达。
关键词： profilin 基因；3′RACE ；5′RACE;油菜花粉；RT-PCR 技术
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