[Construction of plant expression plasmid of chimera SBR-CT delta A1].
OBJECTIVE
The purpose of this study is to construct plant expression plasmid containing the gene encoding chimera SBR-CT delta A1.


METHODS
The target gene fragment P2, including the gene-encoded chimera SBR-CT delta A1 (3,498-5,378 bp), was obtained by standard PCR amplification. The PCR products were ligated with pGEM-easy vector through TA clone to form plasmid pTSC. The plasmid pTSC and plasmid pPOKII were digested by restricted endonuclease BamHI and KpnI, and the digested products were extracted and purified for recombination. Then the purified P2 and plasmid pPOKII were recombined by T4 DNA ligase to form recombinant plasmid pROSC; inserting bar gene into the plasmid and form pROSB plasmid. The recombined plasmids were isolated and identified by restricted endonuclease cutting and Sanger dideoxy DNA sequencing.


RESULTS
P2 gene was linked to pPOKII plasmid and formed recombinant plasmid pROSC. The DNA sequence and orientation were corrected. And bar gene was inserted into pPOSC and form recombinant plasmid pROSB.


CONCLUSIONS
Plant expression vector pROSC and pROSB containing the gene encoding chimera SBR-CT delta A1, which may provide useful experiment foundation for further study on edible vaccine against caries have been successfully constructed.
D018829:Adhesins, Bacterial / Q000235:genetics; D000818:Animals; D002678:Chimera / Q000235:genetics; D003001:Cloning, Molecular; D003731:Dental Caries / Q000517:prevention & control; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008074:Lipoproteins / Q000235:genetics; D018551:Solanum lycopersicum / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic; D022541:Streptococcal Vaccines / Q000235:genetics* / Q000276:immunology; D013295:Streptococcus mutans / Q000235:genetics* / Q000276:immunology; D019444:Vaccines, DNA / Q000235:genetics*
D018829:Adhesins, Bacterial; D008074:Lipoproteins; D011994:Recombinant Proteins; D022541:Streptococcal Vaccines; D019444:Vaccines, DNA; C067560:saliva-binding protein, Streptococcus sanguis 12