病程相关蛋白(Pathogenesis-related proteins,PRs)是植物受生物或非生物胁迫后诱导并积累的一类蛋白质,具有抗病抗逆的多种功能。为检测本生烟病程相关蛋白10(Nicotiana benthamiana Pathogenesis-related protein 10,NbPR10)的表达,...
详细信息
病程相关蛋白(Pathogenesis-related proteins,PRs)是植物受生物或非生物胁迫后诱导并积累的一类蛋白质,具有抗病抗逆的多种功能。为检测本生烟病程相关蛋白10(Nicotiana benthamiana Pathogenesis-related protein 10,NbPR10)的表达,制备其抗血清,并用以探究其可能的特性和功能。将已有的NbPR10基因构建到原核表达载体p GEX-KG中获得重组质粒p GEXKG-NbPR10,转化大肠杆菌BL21(DE3),经0.2 m M IPTG诱导表达分子量约44 k Da的融合蛋白(含GST标签),用GST亲和柱纯化获得纯化的融合蛋白。以纯化的融合蛋白为抗原免疫家兔制备NbPR10蛋白的抗血清,经Western blot测定抗血清效价达1∶10 000,能至多检测到稀释比例为1∶320的转基因本生烟叶片中的PR10蛋白或1 ng原核表达的蛋白,且能检测到本生烟根和茎中的PR10蛋白,证明PR10在本生烟根和茎中表达量较在叶部高,为进一步研究NbPR10蛋白的功能提供参考。
The 3’ terminal genomic 1 611 nucleotides of Beet mosaic virus Xinjiang isolate(BtMV-XJ)from China was determined(GenBank accession number DQ345522).The sequence started within a single open reading frame which was e...
详细信息
The 3’ terminal genomic 1 611 nucleotides of Beet mosaic virus Xinjiang isolate(BtMV-XJ)from China was determined(GenBank accession number DQ345522).The sequence started within a single open reading frame which was expected to encode part C-terminus of NIb protein of 201 amino acids,complete capsid protein(CP)of 276 amino acids and followed by a 3’ untranslated region(UTR)of 168 *** comparison of sequence diversity among BtMV-XJ and other three previously reported isolates revealed that it shared 98.3%,98.1%,91.4% nucleotide identity,and 99.79%,99.58%,97.69% deduced aa sequence identity with the corresponding 3’ terminal regions of BtMV-SL,BtMV-UK and BtMV-US,respectively,and that most nucleotide mutations were silent with no effect on aa *** addition,a virus-specific and ra-pid RT-PCR detection method of BtMV was also developed based on the nucleotide sequence obtained.
暂无评论