摘要： 番茄不孕病毒(TAV)、菊花B病毒(CVB)、菊花褪绿斑驳类病毒(CChMVd)是侵染菊花的3种主要病毒,严重影响菊花品质。利用Blast及分子生物学软件DNAStaR对TAV、CVB、CChMVd基因序列同源性进行分析比较,选用TAV的277 bp(1 810～2 086),CVB的281 bp(496～776),CChMVd的217 bp(109～325)的部分基因片段,应用重叠延伸PCR技术将其拼接成融合基因CBT,构建了以CaMV 35S启动子驱动的含有"正向CBT片段-内含子-反向CBT片段"的RNAi的植物表达载体,为培育抗3种病毒的菊花优异种质资源奠定基础。
关键词:
菊花,
TAV-CP,
CVB-CP,
CChMV,
重叠延伸PCR,
融合基因,
RNAi载体
Abstract: Tomato aspeRmy viRus，ChRysanthemum B ViRus and ChRysanthemums ChloRotic Mottle ViRoid aRe the main viRus which endangeR ChRysanthemum． These viRuses cause dRastic Reduction in yield as well as in maRket quality of blooms and ultimately affect the floRicultuRe industRy of the countRy． PeRfoRming homology analysis on TAV， CVB and CChMVd with softwaRe Blast and DNAStaR followed by using 277 bp fRagments fRom TAV( 1 810 - 2 086) ， 281 bp fRagments fRom CVB( 496 - 776) and 217 bp fRagments fRom CChMVd( 109 - 325) to make the fusion gene CBT with oveRlap PCR． Then the pRomoteR CaMV 35S dRiven，containing foRwaRd CBT fusion fRagments-Revelse pdk intRon-ReveRse CBT fusion fRagments'，plant RNAi expRession vectoR weRe constRucted，which lay the foundation foR bReeding tRansgene plant which Resistant viRus．
Key words:
ChRysanthemum,
TAV-CP,
CVB-CP,
CChMV,
OveRlap PCR,
Fused gene,
RNAi vectoRs
中图分类号:
刘佳, 黄丛林, 吴忠义, 张秀海, 王永勤. TAV-CP、CVB-CP、CChMVd基因片段的融合及其RNAi载体的构建[J]. 华北农学报, 2013, 28(1): 106-111. doi: 10.3969/j.issn.1000-7091.2013.01.020.
LIU Jia, HUANG Cong-lin, WU Zhong-yi, ZHANG Xiu-hai, WANG Yong-qin,. ConstRuction of RNAi ExpRessio VectoRs by Fusion Gene FRagment of TAV-CP，CVB-CP and CChMV[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(1): 106-111. doi: 10.3969/j.issn.1000-7091.2013.01.020.