| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | A 3’ RACE PCR was performed using a hotstart Taq DNA polymerase (Takara, Japan). PCR products were collected and inserted into pMD19-T vectors (Takara, Japan), then custom sequenced (Genscript, China). Reverse primers used for 5’ RACE were designed based on the sequencing results (Table 1: Beta-tub reverse outer and Beta-tub reverse inner) and the 5’ RACE PCR was done following the manufacturer’s instructions (Takara, Japan). | Get A Quote |
Marssonina brunnea is the causal agent of Marssonina leaf spot of poplar. We successfully transformed M. brunnea using Agrobacterium tumefaciens. A highly-efficient promoter was a key factor in the construction of this transformation system construction. The beta -tubulin gene is stable and highly expressed at almost all stages of the life history of M. brunnea. A β-tubulin gene promoter was obtained based on a genomic sequence using an adapter ligation-mediated PCR. This was then inserted into the pMDC83 plasmid vector for high expression of the inserted gene. A genomic DNA PCR assay and fluorescence observation of transformants authenticated successful transformation and fluorescence intensity indicated the ... More
