[Construction of plant expression vectors containing the gene encoding cholera toxin B subunit].
OBJECTIVE
To construct the plant expression vector containing the nucleotide sequence encoding cholera toxin B (CTB) subunits.


METHODS
Using high-fidelity PCR, we amplified CTB genes that were then subcloned into the transition vector pRTL2. Following confirmation of the CTB nucleotide sequence, the vector was subcloned into the plant vector pBI121 that was subsequently transferred into Agrobacterium tumefaciens LBA4404 by electroporation.


RESULTS
CTB DNA that was ligated into the transition vectors resulted in the 2 vectors designated as pRCTB and pRCTBK. After the 2 vectors were ligated into the plant binary vector pBI121 respectively, new plant binary vectors, namely pBI-CTB and pBI-CTBK, were produced. Analysis with restriction endonucleases confirmed successful transfer of pBI-CTB and pBI-CTBK into Agrobacterium tumefaciens LBA4404.


CONCLUSIONS
With appropriate technological strategy, the plant binary expression vectors encoding CTB have been constructed, which facilitates further investigation of CTB protein expressions in transgenic plant.
Peng
Zhi-Qiang
ZQ
Department of Epidemiology, First Military Medical University, Guangzhou 510515, China.
Yu
Shou-Yi
SY
Yu
Di-Qiu
DQ
He
Zhu-Mei
ZM
D002772:Cholera Toxin / Q000096:biosynthesis / Q000235:genetics*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017343:Genes, Plant; D005822:Genetic Vectors / Q000235:genetics
D002772:Cholera Toxin