以高丛蓝莓(Highbush blueberry)"喜来(Sierra)"和"日出(Sunrise)"为研究材料,通过RACE(rapid amplification of c DNA end,RACE)技术获得蓝莓(Vaccinium ssp.)F3′H基因的全长c DNA序列,并使用生物信息学工具对该...以高丛蓝莓(Highbush blueberry)"喜来(Sierra)"和"日出(Sunrise)"为研究材料,通过RACE(rapid amplification of c DNA end,RACE)技术获得蓝莓(Vaccinium ssp.)F3′H基因的全长c DNA序列,并使用生物信息学工具对该基因编码蛋白的氨基酸组成及理化性质、二级和三级结构,以及氨基酸序列的相似性等进行了预测和分析。利用荧光定量PCR(RT-q PCR)手段来检测其在蓝莓果实不同发育时期表达情况。结果表明:Vs F3′H全长c DNA为1 871 bp,编码530个氨基酸,蛋白相对分子质量为58.33 k D,理论p I值为7.32;蛋白疏水性指数为0.004,属于疏水性蛋白,二三级结构预测可知其富含α-螺旋和无规则卷曲。RT-q PCR发现Vs F3′H主要在果实发育前期表达量较高,蓝果期表达量明显下调,且在不同品种中以蓝果期的表达量差异显著。这些研究结果为解析蓝莓果实色泽发育及果树分子育种奠定了理论基础。展开更多
One resistance gene analog fragment named RGA-CIN14 was isolated from TcLr19 wheat,which contains kinase-2,kinase-3a,and the GLPL motif of the NBS-spanning region,using degenerated primers according to the nucleotide ...One resistance gene analog fragment named RGA-CIN14 was isolated from TcLr19 wheat,which contains kinase-2,kinase-3a,and the GLPL motif of the NBS-spanning region,using degenerated primers according to the nucleotide binding site （NBS） conserved domain.Based on the RGA-CIN14,a full-length cDNA,CIN14,which was 2 987 bp encoding 880 amino acids,was obtained by using the method of the rapid amplification cDNA ends （RACE）.Bioinformatics analysis showed that the deduced amino acids of CIN14 protein consisted of a NB-ARC conserved domain and many leucine-rich repeats （LRR） domains.The phylogenetic tree analysis indicated a considerable identity of the protein encoded by CIN14 with that of wheat leaf rust resistance gene Lr1,but a lower similarity with Lr21.The expression profile of the CIN14 gene detected by semi-quantitative RT-PCR showed that the CIN14 gene was not induced by Puccinia triticina and it was a constitutive gene with low abundance in the wheat leaf tissue.The resistance homology sequence was successfully obtained,which provides the shortcut for cloning of the resistance gene in TcLr19 wheat.展开更多
文摘以高丛蓝莓(Highbush blueberry)"喜来(Sierra)"和"日出(Sunrise)"为研究材料,通过RACE(rapid amplification of c DNA end,RACE)技术获得蓝莓(Vaccinium ssp.)F3′H基因的全长c DNA序列,并使用生物信息学工具对该基因编码蛋白的氨基酸组成及理化性质、二级和三级结构,以及氨基酸序列的相似性等进行了预测和分析。利用荧光定量PCR(RT-q PCR)手段来检测其在蓝莓果实不同发育时期表达情况。结果表明:Vs F3′H全长c DNA为1 871 bp,编码530个氨基酸,蛋白相对分子质量为58.33 k D,理论p I值为7.32;蛋白疏水性指数为0.004,属于疏水性蛋白,二三级结构预测可知其富含α-螺旋和无规则卷曲。RT-q PCR发现Vs F3′H主要在果实发育前期表达量较高,蓝果期表达量明显下调,且在不同品种中以蓝果期的表达量差异显著。这些研究结果为解析蓝莓果实色泽发育及果树分子育种奠定了理论基础。
基金funded by the National Natural Science Foundation of China (30771391,30700505)the Natural Science Foundation of Hebei Province,China (C2008000281)
文摘One resistance gene analog fragment named RGA-CIN14 was isolated from TcLr19 wheat,which contains kinase-2,kinase-3a,and the GLPL motif of the NBS-spanning region,using degenerated primers according to the nucleotide binding site （NBS） conserved domain.Based on the RGA-CIN14,a full-length cDNA,CIN14,which was 2 987 bp encoding 880 amino acids,was obtained by using the method of the rapid amplification cDNA ends （RACE）.Bioinformatics analysis showed that the deduced amino acids of CIN14 protein consisted of a NB-ARC conserved domain and many leucine-rich repeats （LRR） domains.The phylogenetic tree analysis indicated a considerable identity of the protein encoded by CIN14 with that of wheat leaf rust resistance gene Lr1,but a lower similarity with Lr21.The expression profile of the CIN14 gene detected by semi-quantitative RT-PCR showed that the CIN14 gene was not induced by Puccinia triticina and it was a constitutive gene with low abundance in the wheat leaf tissue.The resistance homology sequence was successfully obtained,which provides the shortcut for cloning of the resistance gene in TcLr19 wheat.
