麻疯树JcCAULIFLOWER A-Like基因的克隆、原核表达及表达模式分析

Cloning, prokaryotic expression and expression pattern analysis of Jatropha curcas JcCAULIFLOWER A-Like gene

CAULIFLOWER(CAL)是与花分生组织的形成有关的基因。通过RACE-PCR技术从麻疯树花蕾中成功获得了1个CAL(JcCAL A-Like)基因c DNA的全长序列,并对其原核表达产物进行质谱鉴定,分析了其亚细胞定位情况及其在麻疯树花不同发育时期的表达情况。结果表明:JcCAL A-Like cDNA全长1021 bp,开放阅读框长603 bp,编码200个氨基酸,蛋白质的大小约为23.368 kDa,等电点pI为9.63;表达情况提示该基因可能参与了麻疯树雄花单核花粉粒的形成及雌花的胚囊成熟;此外,亚细胞定位分析显示该基因定位于细胞核中,这表明JcCAL A-Like发挥功能的场所是在细胞核。

The CAULIFLOWER(CAL)is thought to be involved in the formation of floral meristems.In this study,the full-length cDNA of a CAL,named as JcCAL A-Like,was successfully obtained from Jatropha curcas using RACE-PCR.The prokaryotic expression product of this gene was identified by using mass spectrometry.Its subcellular localization was analyzed by employing transient expression system.The expression of JcCAL A-Like in Jatropha curcas flower at different developmental stages was analyzed by qRT-PCR.The results showed JcCAL A-Like cDNA was 1021 bp,its open reading frame was 603 bp which encoded a protein with 200 amino acids.This gene encoded a mass of 23.368 kDa and a pI of 9.63 protein.The expression levels in Jatropha curcas indicated that this gene might be involved in the formation of mononuclear pollen grains and maturity of female flowers.In additional,subcellular localization analysis indicated that JcCAL A-Like would function in nucleus.