甘蓝型油菜小核糖核蛋白BnSmD1编码区全长cDNA的克隆与表达分析  

Identification and expression analysis of a full-length cDNA encoding Brassica napus small nuclear ribonucleoprotein BnSmD1

作　　者：袁晓萌[1] 周云涛[1] 张红岩[1] 薛华[1] 周琳[1] 赵云[1] 

机构地区：[1]四川大学生命科学学院教育部生态环境与资源重点实验室,成都610064

出　　处：《遗传》2007年第12期1525-1528,共4页Hereditas（Beijing)

基　　金：国家自然科学基金(编号:30571174)资助项目~~

摘　　要：通过筛选野生型油菜(Pet33-10)与无花瓣油菜(Apet33-10)反向消减文库(SSH)和运用RACE-PCR技术,获得了甘蓝型油菜小核糖核蛋白BnSmD1的全长编码区cDNA(GenBank登陆号DQ298446)。该基因长484 bp,含有一个长354 bp的阅读框。BnSmD1在N端拥有两个高度保守的结构域(Sm-1和Sm-2),羧基端则含有一个RG重复序列。Northern blot表达结果显示:BnSmD1在甘蓝型油菜的各个组织均有表达,但是它在早期花蕾中的表达明显高于同期的叶和茎。通过对BnSmD1在Apet33-10无花瓣品系与野生型有花瓣品系Pet33-10中各组织的表达差异进行比较,发现该基因在Apet33-10的早期花蕾中表达明显下降。因此,BnSmD1可能对植物的早期花发育起到了重要的作用,并很有可能影响花瓣的形成。By using substractive hybridization （SSH） and rapid amplification of cDNA ends-polymerase chain reaction （RACE-PCR）, a full-length cDNA encoding Brassica napus small nuclear ribonucleoprotein, named BnSmD1, was obtained. It had 484 base pairs in length containing an open reading flame （ORF） of 354 bp and encoding a predicted protein of 118 amino acids with a molecular weight of 13 kDa. The BnSmD1 protein shares two highly conserved Sm folds （Sm-1 and Sm-2） and a C-terminal RG dipeptide repeat. Northern blot analysis revealed that BnSmD1 was expressed in all tested organs in B. napus, but its transcript level in early floral buds was much higher than that in leaf and stem tissues. No obvi- ous expression difference was observed in leaf and stem tissues between the apetalous line Apet33-10 petalled near-isogenic line Pet33-10. Compared with wild type, the expression of BnSmD1 in the early floral buds of apetalous mutant Apet33-10 was significantly reduced. Taken together, our results suggest that BnSmD1 may play an important role in early floral petal development in B. napus.

关 键 词：甘蓝型油菜 早期花发育 Northern BLOT RT-PCR 抑制性消减杂交 

分 类 号：Q943.2[生物学—植物学]