By the previous studies, it was reported that the TMV gene exsists only in cytoplasm and the splicing of pre-mRNA, which occures in nuleus, does not happen in cytoplasm. But in this study, we found that the expression of GUS harbored in vector of TMV coincided with intron splicing of pre-mRNA.
In an attempt to use TMV vector to express a fusion protein in Nicotiana benthenmiana , a GUS gene with an intron was cloned into the vector, Supprisingly, the GUS gene was expressed at high level. Reverse-transcription and polymerase chain reaction(RT-PCR), DNA sequencing and GUS-INTRON gene mutant analyses of transcripts show an event ofintron splicing happened with the GUS gene.
Ye Xinfu,, Quinn Li
. Pre-mRNA Splicing of Genes in a TMV Viral Vector[J]. Chinese Agricultural Science Bulletin, 2005
, 21(2)
: 31
-31
.
DOI: 10.11924/j.issn.1000-6850.050231
1Creager,AN,Scholthof,KB,Citovsky,VV,Scholthof,HB. Tobacco mosaic virus. Pioneering research for a century[J].Plant Cell,1999,11:301~8
2Dawson W O, Lehto K M.. Regulation of tobacco virus gene expression[J].Adv Virus Res, 1990,38:307~42
3Turpen T H, Dawson W O. Amplification, movement and expression of genes in plants by viral based vectors. In: Transgenic Plants:Fundamentals and Applications,ed.A.Hiatt. 1992.195~217..
4Meeusen R. Plant Biotechnology: beyond the beach head .In International Molecular Farming Conference.pp.W4.London,Ontario,Canada:Agriculture and Agri-food Canada. 1999.
5Hills G J, Plaskitt K A, Young N D, et al. Immunogold localization of the intracellular sites of structural and nonstructural tobacco mosaic virus proteins[J].Virology,1987,161:488~96
6Okamoto S, Machida Y, Takebe I. Subcellular localization of tobacoo mosaic virus minus strand RNA in infected protoplasts[J].Virology,1988,167:194~200
7吴乃虎.基因工程原理(下)[S].北京:科学出版社,2002.65~88
8 Beven F, Simpson G G, Brown J W, et al. The organization of spliceosomal components in the nuclei of higher plants[J]. J cell Sci,1955.108:509~18
9Sleeman J E, Lamond A I. Nuclear organization of pre-mRNA splicing factors[J].Curr Opin Cell Biol, 1999,11:372~7
10 Lopez-Moya J J, Garcia J A.. Construction of a highly infectious intron-containing cDNA clone of plun pox potyvirus and its use to infect plants by particle bombardment[J].Virus Res,2000,68:99~107
11Simposon G G, Filipowicz W. Splicing of precursors to mRNA in higher plants: mechanism, regulation and sub-nuclear organization of the spliceosomal machinery[J].Plant Mol Biol,1996,32:1~41
12Connor-Sanchez A, Willmitzer L, Rocha-Sosa M. Construction of an introncontaining marker gene: splicing of the intron in transgenic plants and its use in monitoring early events in Agrobacterium-mediated plant trasformation[J].Mol Gen Genet,1990,220:245~50
