根据已知的大麦黄矮病毒GPV株系的外壳蛋白(Coat Protein CP)和移动蛋白(Movement Protein MP)基因序列合成了CP、MP基因的上下游引物,通过PCR扩增获得含有目的基因的片段,经过Sal I和Pst I双酶切、连接、转化、重组质粒的酶切鉴定及基...根据已知的大麦黄矮病毒GPV株系的外壳蛋白(Coat Protein CP)和移动蛋白(Movement Protein MP)基因序列合成了CP、MP基因的上下游引物,通过PCR扩增获得含有目的基因的片段,经过Sal I和Pst I双酶切、连接、转化、重组质粒的酶切鉴定及基因测序,构建了酵母表达载体pGBKT7-GPV-CP和pGBKT7-GPV-MP,用于在酵母双杂交分析中表达诱饵融合蛋白,为进一步筛选小麦cDNA文库内与大麦黄矮病毒互作的寄主因子、克隆寄主因子、推测其种类和功能奠定了基础。展开更多
Young leaves and shoot with Apple stem pitting virus（ASPV） detected by RT-PCR（reverse transcription-polymerase chain reaction） from Korla pear were used as materials and total RNA was extracted.Two expected fragme...Young leaves and shoot with Apple stem pitting virus（ASPV） detected by RT-PCR（reverse transcription-polymerase chain reaction） from Korla pear were used as materials and total RNA was extracted.Two expected fragments had been obtained by RT-PCR with two specific pairs of primers based on the sequence of NC003462 published in GenBank.The specific fragments had been recovered and cloned,and the white colonies had been screened out.After enzyme digestion of the plasmids isolated,the positive clone was selected to be sequenced.Compared with NC003462,three ORFs（open reading frame）-ORF2,ORF3 and ORF4 of ASPV with 673 bp,365 bp,276 bp representing homology of 77%,88.52%,86.33% respectively had been obtained.展开更多
文摘根据已知的大麦黄矮病毒GPV株系的外壳蛋白(Coat Protein CP)和移动蛋白(Movement Protein MP)基因序列合成了CP、MP基因的上下游引物,通过PCR扩增获得含有目的基因的片段,经过Sal I和Pst I双酶切、连接、转化、重组质粒的酶切鉴定及基因测序,构建了酵母表达载体pGBKT7-GPV-CP和pGBKT7-GPV-MP,用于在酵母双杂交分析中表达诱饵融合蛋白,为进一步筛选小麦cDNA文库内与大麦黄矮病毒互作的寄主因子、克隆寄主因子、推测其种类和功能奠定了基础。
文摘Young leaves and shoot with Apple stem pitting virus（ASPV） detected by RT-PCR（reverse transcription-polymerase chain reaction） from Korla pear were used as materials and total RNA was extracted.Two expected fragments had been obtained by RT-PCR with two specific pairs of primers based on the sequence of NC003462 published in GenBank.The specific fragments had been recovered and cloned,and the white colonies had been screened out.After enzyme digestion of the plasmids isolated,the positive clone was selected to be sequenced.Compared with NC003462,three ORFs（open reading frame）-ORF2,ORF3 and ORF4 of ASPV with 673 bp,365 bp,276 bp representing homology of 77%,88.52%,86.33% respectively had been obtained.
