甘蓝型油菜生物节律钟输出基因BnGI的克隆和表达分析

利用同源克隆的方法,以拟南芥GIGANTEA基因为种子序列,在甘蓝型油菜中克隆得到GI基因,命名为BnGI。序列分析结果表明:该基因编码区全长为3 516bp,编码1 171个氨基酸,与白菜GI基因序列同源性达99%,与拟南芥GI基因同源性达87%。成熟蛋白的等电点为6.59,分子量为127.15kDa,亚细胞定位预测在细胞核定位。荧光半定量检测结果表明:BnGI基因在根、茎、顶端分生组织、叶、花等中均有表达,但表达水平具有组织特异性。

Cloning and Expression Analysis of Circadian Clock Output Gene BnGI in Brassica napus L.

A cDNA fragment of GIGANTEA gene was cloned from Brassica napus L according to the method of homologous cloning with the sequence of Arabidopsis thaliana GI as seeding sequence.Sequence analysis showed that the full length of this gene cDNA coding region was 3 516 bp encoding 1 171 amino acids.It had 99％ homologies with sequence of Brassica rapa GI and 87 ％ homologies with sequence of Arabidopsis thaliana GI.The mature protein isoelectric point was at pH8.47 and molecular weight was 127.15 kDa.Furthermore,the protein was predicted predominantly located in the nucleus.Fluorescence semi-quantitative PCR analysis indicated that BnGI was expressed in all organs of the plant,including roots,stems,apical meristem,leaves and flowers.However,the expression level of BnGI was tissue-specific.