Construction of a Plant Inducible Expression Vector and Its Transient Expression in Tobacco

doi:10.3969/j.issn.1000-7091.2013.04.008

Abstract

Abstract: According to the sequences of pathogen- inducible plant promoters PPP3 at GenBank，PPP3 promot- ers was cloned from tobacco genome．It was used to replace cauliflower mosaic virus 35S promoter of pCAM- BIA1301．The recombinant plasmid was used to transform Agrobacterium tumefaciens GV3101．The inducibility of the PPP3 promoters in tobacco leaf was evaluated by Agrobacterium tumefaciens- transient genetic transformation as- sye．Real- time quantitative PCR was used to screen the PPP3 promoters with high inducible expression．The results showed that the gus transcript level under the control of PPP3 promoters increased respectively 27．94 fold and 17. 69 fold after inoculation with Ralstonia solanacearum and SA．The PPP3 promoter had the advantages such as low basal activity and high expression activity．

Key words: Inducible expression Promoter, Transient expression, Real- time quantitative PCR

CLC Number:

Q786

OUYANG Le-jun, HUANG Zhen-chi, SHA Yue-e, ZENG Fu-hua. Construction of a Plant Inducible Expression Vector and Its Transient Expression in Tobacco[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(4): 41-45. doi: 10.3969/j.issn.1000-7091.2013.04.008.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.3969/j.issn.1000-7091.2013.04.008

http://www.hbnxb.net/EN/Y2013/V28/I4/41

References

[1] 邱礽，陶刚，李奇科，等农杆菌渗入法介导的基因瞬时表达技术及应用[J]. 分子植物育种，2009，7 (5): 1032-1039．
[2] Yang Y，Li R，Qi M． In vivo analysis of plant promoters and transcription factors by agroinfiltration of tobacco leaves[J]. Plant J，2000，22(6): 543-551．
[3] Negrouk V． Highly efficient transient expression of func-tional recombinant antibodies in lettuce[J]. Plant Sci-ence，2005，169: 433-438．
[4] Srinivas L． Transient and stable expression of hepatitis B surface antigen in tomato (Lycopersicon esculentum L． ) [J]. Plant Biotechnol Rep，2008，2: 1-6．
[5] Santos-Rosa M． Development of a transient expression sys-tem in grapevine via agroinfiltration[J]. Plant Cell Rep，2008，27: 1053-1063．
[6] Choi J J． A promoter from pea gene DRR206 is suitable to regulate an elicitor-coding gene and develop disease re-sistance[J]. Phytopathology，2004，94: 651-660．
[7] Hoffmann T． RNAi-induced silencing of gene expression in strawberry fruit (Fra-garia ananassa) by agroinfiltra-tion: a rapid assay for genefunction analysis[J]. Plant J，2006，48: 818-826．
[8] 彭舒，黄真池，欧阳乐军，等．植物基因工程中人工启动子的研究进展[J] . 植物生理报，2011，47(2): 141-146．
[9] Rushton P J． Synthetic plant promoters containing defined regulatory elements provide novel insights into pathogen-and wound-induced signaling[J]. Plant Cell，2002，14: 749-762．
[10] 魏开发．水仙胚性愈伤的获得及农杆菌介导 GUS 基因的遗传转化[J]. 南京林业大学学报: 自然科学版，2009，33(4): 33-37．
[11] Pfaffl M W． Relative expression software tool(REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR[J]. Nucl Acids Res，2002，30: 249-259．
[12] Peng J L，Bao Z L，Li P，et al．，Hanpin and its functional domains activate pathogen-inducible plant promoters in Arabidopsi[J]，Acta Botanica Sinica，2004，46: 1083-1090．

Please choose a citation manager

Content to export