摘要
将来自粟酒裂殖酵母的pac1基因,导入原核表达载体pET-5a中,并转入大肠杆菌BL21(DE3)pLysS,经IPTG诱导,其表达产物能够降解4种植物病毒Cucumber mosaic virus(CMV)、Tobacco mosaic virus(TMV)、Rice black-streakeddwarf(RBSDV)和Rice dwarf virus(RDV)和3种类病毒Apple scar skin viroid(ASSVd)、Coleus blumei viroid(CBVd)和Hopstunt viroid(HSVd)的dsRNA。将pac1导入双元载体pBI121,并转入根癌农杆菌LBA4404,以烟草(品种NC89)组培苗的叶片为受体材料进行转化,经过诱导愈伤、分化、再生和筛选培养,获得了50株Kan抗性植株,收获T1种子分别播种,对这些转基因植株进行分子生物学检测。PCR、PCR-Southern和RT-PCR检测结果表明,pac1基因已整合到受体基因组中。
pac 1 gene was obtained from Schizosaccharomyces pombe and transferred into pET-5a expression system.The expression product of cloned pac 1 in Escherichia coli BL21（DE3）pLysS which induced by the IPTG showed activity to digest dsRNAs of 4 plant viruses Cucumber mosaic virus（CMV）,Tobacco mosaic virus（TMV）,Rice black-streaked dwarf（RBSDV）,Rice dwarf virus（RDV）and 3 viroids Apple scar skin viroid（ASSVd）,Coleus blumei viroid（CBVd）,Hop stunt viroid（HSVd）.Harboring the binary vector pBI121,which contained pac 1 gene,Agrobacterium tumefaciens strain LBA4404 was used to transform the pre-cultured tobacco leaves.After callus induction,differentiation,regeneration and selection culture stage,totally 50 Kan resistant plants were obtained,then T1 seeds were planted separately.PCR,PCR-Southern and RT-PCR detection of the transgenic plants revealed the integration of the pac 1 gene in the tobacco genomes.
出处
《植物病理学报》
CAS
CSCD
北大核心
2008年第5期489-495,共7页
Acta Phytopathologica Sinica
基金
国家自然科学基金资助项目(30571260
30771403)
