摘要
为研究甘蓝型油菜磷酸甘油酸激酶(PGK)基因表达特性,在对拟南芥PGK基因家族生物信息学分析的基础上,通过电子克隆方法获得3个甘蓝型油菜PGK基因(BnPGK1、BnPGK2、BnPGK3)。分别设计特异引物,以甘蓝型油菜雄性不育系09A和保持系09B的cDNA为模板克隆BnPGK基因全长序列。根据获得的cDNA序列设计实时荧光定量特异引物,采用实时荧光定量PCR技术,研究油菜雄性不育系与保持系PGK基因表达差异。结果显示:BnPGK基因在甘蓝型油菜雄性不育系09A和保持系09B的根、茎、叶、花蕾中均有表达,属组成性表达。除茎中的BnPGK3外,BnPGK其它基因在根、茎、叶中的表达均表现为09A高于09B,而在花蕾中均为09B高于09A,BnPGK1和BnPGK3在09B中的表达量是09A中的2倍以上。
Three full-length cDNA sequences of PGK genes（BnPGK1,BnPGK2 and BnPGK3）were obtained through electronic cloning method based on bioinformatics analysis of Arabidopsis PGK gene family for the study of Phosphoglycerate kinase（PGK）gene expression characteristics fromBrassica napus.Specific primers were designed and cDNA of male sterile line 09 Aand maintainer line 09 Bwere as templates to clone BnPGKfull-length sequences.Real-time fluorescence quantitative specific primers were designed according to the obtained cDNA sequences.The Real-time quantitative analysis was conducted by using Quantitative PCR method.Results showed that the BnPGK genes expressed variously in different tissues,such as root,stem,leaf and bud of 09 Aand 09B,belong to constitutively expressed.Except BnPGK3 in the stem,the expression of BnPGK other genes of 09 Awere higher than that of 09 Bin root,stem and leaf,but the expression of BnPGK genes of 09 Bwere higher than that of 09 Aand the expression of BnPGK1 and BnPGK3of 09 Bwas 2times than that of 09 Ain bud.
出处
《西北植物学报》
CAS
CSCD
北大核心
2014年第11期2188-2193,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
国家自然科学基金(31171588)
重庆市自然科学基金(cstc2012jjA80010)
