Construction Of A Plant Expression Vector Based On Turnip Mosaic Virus Isolate CDN1 And Cloning Of The CDNA Of Turnip Mosaic Virus Isolate CHN5

Turnip mosaic virus (TuMV) is one of the most important pathogens on cruciferae crops. A new DNA sequence coding the recognition site of NIa protease on polyprotein was designed based on the analysis of the NIa protease recognition sites on TuMV different isolates. A multi-clone site(MCS) was designed based on the analysis of the conservative absent restriction sites on TuMV different isolates. The TuMV isolate CDN1 infectious full-length cDNA clone pVIR95T was modified into an expression vector pVIR95TM by inserting the adaptor with the MCS and the new NIa recognition site after the site between the Nib and CP by PCR. The DNA sequence coding the human a -calcitonin gene related peptide(hCGRP) was acquired by DNA synthesis and PCR method, and was inserted into the MCS of pVIR95TM to form an expression vector expressing the hCGRP on its host plant.According to the conservative sequence of TuMV different isolates, a set of primers was desinged to amplify the 9. 4kb cDNA of TuMV isolate CHN5 covering about 94% of its full genome by immunocapture RT-PCR, and cloned the cDNA into a T-vector. 8.45Kb part of cDNA of TuMV isolate CHN5 substitued for the corresponding cDNA of TuMV isolate CDN1 on pVIR95TM to form TuMV isolate CHN5 infectious cDNA clone.