As a critical enzyme in secondary metabolism of plant,Phenylanlanine ammonialyase(PAL) has significant meaning to its development and strong tolerance ability against hard.A full-length cDNA of PAL gene was isolated f...As a critical enzyme in secondary metabolism of plant,Phenylanlanine ammonialyase(PAL) has significant meaning to its development and strong tolerance ability against hard.A full-length cDNA of PAL gene was isolated from Phyllostachys edulis through RT-PCR and RACE methods,and named as PePAL1(GenBank accession number:FJ195650).PePAL1 is 2 436 bp,which contains an open reading frame encoding 701 amino acids.The results of amino acid sequence analysis showed that PePAL1 had high identities with other gramineous PAL in Oryza sativa,Zea mays,Saccharum officinarum,Bambusa ventricosa,Bambusa oldhamii and Triticum aestivum,especially with PAL from O.sativa up to 93.0%.Phylogenetic analysis showed that PePAL1 and LLB1 were on different branch sites.Tissue specific expression showed that PePAL1 expressed in leaf,sheath,stem and root,much higher in stem.展开更多
采用RT-PCR方法从小麦品种豫教2号发育籽粒中克隆出小麦淀粉合成关键酶—AGPase的胞质型小亚基(cytosolic small subunit,SSUⅠ)、质体型小亚基(plastidial small subunit,SSUⅡ)、胞质型大亚基(cy-tosolic large subunit,LSUⅠ)和质体...采用RT-PCR方法从小麦品种豫教2号发育籽粒中克隆出小麦淀粉合成关键酶—AGPase的胞质型小亚基(cytosolic small subunit,SSUⅠ)、质体型小亚基(plastidial small subunit,SSUⅡ)、胞质型大亚基(cy-tosolic large subunit,LSUⅠ)和质体型大亚基(plastidial large subunit,LSUⅡ)的cDNA序列。所克隆的基因cDNA序列长度分别为1 4651、6311、941和535 bp。序列比对结果显示SSUⅠ、LSUⅠ和LSUⅡ与已往报道的大麦、小麦、玉米、水稻等相关基因的同源性较高,而SSUⅡ的cDNA序列为首次克隆,与大麦SSUⅡ相比,5'端缺失编码转移肽的一段序列,表明其可能对质体AGPase活性产生一定影响。通过半定量PCR法发现SSUⅠ与LSUⅠ在籽粒中表达量较高,而SSUⅡ和LSUⅡ在叶片中丰富表达。展开更多
以 p KB、pc DNA 3和 p CPA为原始质粒 ,构建 BL G启动子调控的组织型纤溶酶原激活剂 (t PA)乳腺组织特异性表达载体 p BTA。以 Sal I酶切质粒 p BTA,回收 5 .45 kb基因片段 BL G- t PA- b GHpoly A,通过显微注射 ,导入昆明小鼠受精卵...以 p KB、pc DNA 3和 p CPA为原始质粒 ,构建 BL G启动子调控的组织型纤溶酶原激活剂 (t PA)乳腺组织特异性表达载体 p BTA。以 Sal I酶切质粒 p BTA,回收 5 .45 kb基因片段 BL G- t PA- b GHpoly A,通过显微注射 ,导入昆明小鼠受精卵的雄原核内。共注 136 5枚 ,将存活的 10 5 3枚移植到 5 0只同期发情假孕母鼠的输卵管内。怀孕 2 4只 ,共产仔79只 ,上述仔鼠通过 PCR检测 ,结果 19只阳性。Southern blotting检测上述 19份 PCR阳性小鼠基因组 ,其中 4只为整合阳性 ,说明已获得了乳球蛋白启动子调控下的 t PA转基因小鼠 ,为 t PA在小鼠体内表达研究提供了必要条件。展开更多
文摘As a critical enzyme in secondary metabolism of plant,Phenylanlanine ammonialyase(PAL) has significant meaning to its development and strong tolerance ability against hard.A full-length cDNA of PAL gene was isolated from Phyllostachys edulis through RT-PCR and RACE methods,and named as PePAL1(GenBank accession number:FJ195650).PePAL1 is 2 436 bp,which contains an open reading frame encoding 701 amino acids.The results of amino acid sequence analysis showed that PePAL1 had high identities with other gramineous PAL in Oryza sativa,Zea mays,Saccharum officinarum,Bambusa ventricosa,Bambusa oldhamii and Triticum aestivum,especially with PAL from O.sativa up to 93.0%.Phylogenetic analysis showed that PePAL1 and LLB1 were on different branch sites.Tissue specific expression showed that PePAL1 expressed in leaf,sheath,stem and root,much higher in stem.
文摘采用RT-PCR方法从小麦品种豫教2号发育籽粒中克隆出小麦淀粉合成关键酶—AGPase的胞质型小亚基(cytosolic small subunit,SSUⅠ)、质体型小亚基(plastidial small subunit,SSUⅡ)、胞质型大亚基(cy-tosolic large subunit,LSUⅠ)和质体型大亚基(plastidial large subunit,LSUⅡ)的cDNA序列。所克隆的基因cDNA序列长度分别为1 4651、6311、941和535 bp。序列比对结果显示SSUⅠ、LSUⅠ和LSUⅡ与已往报道的大麦、小麦、玉米、水稻等相关基因的同源性较高,而SSUⅡ的cDNA序列为首次克隆,与大麦SSUⅡ相比,5'端缺失编码转移肽的一段序列,表明其可能对质体AGPase活性产生一定影响。通过半定量PCR法发现SSUⅠ与LSUⅠ在籽粒中表达量较高,而SSUⅡ和LSUⅡ在叶片中丰富表达。
