[Cloning and sequence analysis of tomato fruit-specific E8 promoter from Lycopersicon esculentum (Zhongshu No.5)].
OBJECTIVE
To obtain the gene encoding tomato fruit-specific E8 promoter therefore to prepare for exogenous gene transcription and expression in transgenic tomato fruit.


METHODS
The cotyledons of tomato Lycopersicon esculentum (Zhongshu No.5) were collected for extracting the genomic DNA of this plant. The fruit-specific E81.1 and E82.2 promoter DNA were then amplified by PCR, the product of which was subcloned into pGEM-T vector. After identification by restriction enzymes, the recombinant T-vectors were subjected to sequence analysis.


RESULTS
The fragments of the promoter as amplified by PCR were of predicted length. Digestion with Xba I and Hind III /BamH I proved correct insertion of the target fragments with expected length into the recombinant T vectors. As indicated by homology analysis, the resultant tomato fruit-specific E8 promoter was highly conservative, and E82.2 promoter of Zhongshu No.5, with GenBank submission number of AF515784, proved to share 99% homology with E82.2 promoter of Zhongshu No.5 Cherry as reported by Deikman J.


CONCLUSIONS
Tomato fruit-specific E8 promoter of Zhongshu No.5 has been successfully cloned, thus making possible the subsequent research in oral vaccine of transgenic tomato.
Zhou
Xiao-hong
XH
Department of Parasitology, First Military Medical University, Guangzhou 510515, China. xhzhou2001@sina.com
Chen
Xiao-guang
XG
Zhang
Xiao-dong
XD
Wang
Ya-nan
YN
Li
Lin
L
Xi
Jia-fei
JF
Hu
Jian-jun
JJ
D001483:Base Sequence; D003001:Cloning, Molecular; D018744:DNA, Plant / Q000032:analysis / Q000737:chemistry; D018551:Solanum lycopersicum / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic
D018744:DNA, Plant