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A Seamless Cloning Approach for Porcine Reproductive and Respiratory Syndrome Virus Expression Vector Construction
Published on: May 17, 2024
Construction of the intermediate vector pVBG2307 by incorporating vital elements of expression vectors pBI121 and
1College of Horticulture, Northwest A&F University, Shaanxi, Yangling, P.R. China.
Abstract:
Molecular chaperones of plasmid pBI121 carrying CaMV35S promoter and a nucleotide sequence of plasmid pBI221 were inserted into plasmid pCAMBIA2300 to construct an intermediate vector: pVBG2307. This novel vector pVBG2307 contains a greatly expanded multiple cloning site with an adjacent imported CaMV35S promoter sequence. This vector allows controlled transformation of DNA in both Escherichia coli and Agrobacterium tumefaciens. Cloned PG, orf456, ipt genes and E8, a fruiting promoter, were amplified by PCR of cDNA libraries of Capsicum annum and Lycopersicon esculentum and were then transferred into vector pVBG2307. The viability of this vector was demonstrated, as it regulated PG, orf456, ipt and E8 genes in E. coli and could be transferred into Agrobacterium strain EHA105-4.
