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. 2009 Aug 20;11(1):69–81. doi: 10.1111/j.1364-3703.2009.00574.x

Figure 1.

Figure 1

Construction scheme of pathogenesis‐related protein 10 (PR10) gene silencing vector. The double 35S promoter and the DNA region containing the attR4‐attR3 cassette were amplified by polymerase chain reaction (PCR) separately from their corresponding vectors, and cloned into the corresponding restriction sites in pBluesript II SK‐ vector. The DNA regions corresponding to the PR10 5′arm, intron and 3′ arm were amplified by PCR with primers containing unique homologous recombination sites, cloned into their corresponding entry vectors. A chloramphenicol resistance gene (CmR) selection marker was then inserted into the middle of the PR10 intron before the LR clonase reaction to assemble the RNAi cassette into the pBluescript vector to produce the pBS‐d35S‐attB4‐5′arm‐attB1‐PR10 intron‐CmR‐attB2‐3′arm‐attB3 vector (named pBS‐PR10‐RNAi). The RNAi cassette was then cloned into the pTF102 vector through ligation to produce the final pTF102‐PR10‐RNAi vector.